Arginine and the metabolic regulation of nitric oxide synthesisin cancerRom Keshet and Ayelet Erez
ABSTRACTNitric oxide (NO) is a signaling molecule that plays important roles indiverse biological processes and thus its dysregulation is involved inthe pathogenesis of various disorders In cancer NO has broad andsometimes dichotomous roles it is involved in cancer initiation andprogression but also restricts cancer proliferation and invasion andcontributes to the anti-tumor immune response The importance ofNO in a range of cellular processes is exemplified by its tight spatialand dosage control at multiple levels including via its transcriptionalpost-translational and metabolic regulation In this Review we focuson the regulation of NO via the synthesis and availability of itsprecursor arginine and discuss the implications of this metabolicregulation for cancer biology and therapy Despite the establishedcontribution of NO to cancer pathogenesis the implementation ofNO-related cancer therapeutics remains limited likely due to thechallenge of targeting and inducing its protective functions in a cell-and dosage-specific manner A better understanding of how arginineregulates the production of NO in cancer might thus support thedevelopment of anti-cancer drugs that target this key metabolicpathway and other metabolic pathways involved in NO production
IntroductionNitric oxide (NO) is a short-lived gaseous signaling molecule thatis produced endogenously by a family of enzymes called the nitricoxide synthases (NOS) which catalyze the synthesis of NO fromthe amino acid arginine (Bredt 1999) NO regulates varioussignaling pathways in many different tissues and has diversephysiological roles The most well-known and established functionsof NO relate to its roles in the immune cardiovascular and neuronalsystems Indeed NO is produced by different immune cells mainlymacrophages and is a key regulator of immunity and inflammation(Predonzani et al 2015) It is required for the activation andmigration of macrophages (Connelly et al 2003 Maa et al 2008)and in infectious conditions NO released by immune cells hascytotoxic antimicrobial activities (Woodmansee and Imlay 2003)Conversely NO can serve as an immunosuppressive agent thatlimits T-cell proliferation and activity by promoting apoptosis andby inhibiting cytokine and chemokine production (Bogdan 2015)In the cardiovascular system endothelium-derived NO is a powerfulvasodilator and has a central role in setting vascular tone and blood
pressure (Zhao et al 2015) Moreover NO is known to participatein vascular endothelial growth factor (VEGF)-induced vascularpermeability and angiogenesis (the formation of new blood vessels)(Fraisl 2013) Finally in the nervous system neuronal-derived NOis known to regulate neural development (Kong et al 2014) and toinfluence various brain functions such as cognition and response tostress (Philippu 2016) In the peripheral nervous system NOregulates the function of nerves that regulate smooth-muscle toneand motility in the gastrointestinal tract (Toda and Herman 2005)Thus NO regulates neuronal and blood-vessel functions in mosttissues and as such is essential for the preservation of physiologicalhomeostasis Indeed unregulated NO production is implicated inmultiple pathophysiological conditions including cancer (Burkeet al 2013)
In this Review we summarize what is known about NOmetabolism in carcinogenesis focusing on the importance ofarginine synthesis and its availability for NO productionAdditionally we discuss the role of the arginine-NO axis incancer biology and its potential implications in developing NO-related cancer therapeutics An improved understanding of thismetabolic pathway might enable current treatments to be optimizedand would support the development of other novel anti-cancer drugsthat target tumors in which NO plays a central role in diseaseinitiation and progression
NO synthesis and metabolismIn mammals three distinct genes encode the three NOS isoformsneuronal NOS (nNOS encoded by NOS1) inducible NOS (iNOSencoded by NOS2) and endothelial NOS (eNOS encoded byNOS3) NOS1 and NOS3 are constitutively expressed mainly inneurons and in endothelial cells respectively whereas NOS2 ismainly expressed in immune cells (Mungrue et al 2003) Thebinding of calcium (Ca2+) and calmodulin to nNOS and eNOStransiently activates them to produce nanomolar concentrations ofNO whereas iNOS expression is induced by inflammatorycytokines or by bacterial products such as lipopolysaccharide(LPS) to produce micromolar concentrations of NO (Lind et al2017) Interestingly in addition to cytosolic NOS there is amitochondrial variant of NOS that contributes to the regulation ofNO-related mitochondrial activities (Ghafourifar and Richter1997) All NOS isoforms use arginine as a substrate and requireoxygen NADPH and the cofactor tetrahydrobiopterin (BH4) togenerate NO and citrulline (see Box 1 for a glossary of terms)(Mungrue et al 2003) In this reaction electrons donated byNADPH at the carboxy-terminal reductase domain of NOS arepassed to the heme catalytic center of the oxidase domain whereactivation of molecular oxygen is lsquocoupledrsquo to NO synthesis by twosuccessive mono-oxygenations of arginine NO can also begenerated from the inorganic anions nitrate (NO3
minus) and nitrite(NO2
minus) particularly in hypoxic states (Lundberg et al 2008) Thispathway has been discussed extensively in previous reviews
Department of Biological Regulation Weizmann Institute of Science Rehovot7610001 Israel
Author for correspondence (ayeleterezweizmannacil)
AE 0000-0002-9879-275X
This is an Open Access article distributed under the terms of the Creative Commons AttributionLicense (httpcreativecommonsorglicensesby30) which permits unrestricted usedistribution and reproduction in any medium provided that the original work is properly attributed
1
copy 2018 Published by The Company of Biologists Ltd | Disease Models amp Mechanisms (2018) 11 dmm033332 doi101242dmm033332
Disea
seModelsampMechan
isms
(Lundberg et al 2008) and hence will not be discussed furtherhereCellular NO levels are tightly regulated at several different levels
and by multiple factors NO can be regulated at the level of NOStranscription or via the post-translational modification of NOS aswell as via the cellular expression of the different NOS isoformsand through the availability of NOS substrates such as arginine andBH4 (Mungrue et al 2003) It can also be regulated by the amountof NO produced by the different NOS as well as by the short half-life of NO which is estimated to be in the range of 01ndash2 s andallows the rapid termination of NO signaling cascades once theinitial stimulus is turned off (Thomas et al 2001)The biological effects of NO are exerted through either cyclic
guanosine monophosphate (cGMP) or via post-translationalmodification (PTM) by S-nitrosylation (Box 1) (Seth et al 2018)Canonical NO signaling involves soluble guanylate cyclase (sGC)which is the only known receptor for NO This enzyme is aheterodimer composed of two subunits one ofwhich contains a hemegroup to which NO binds and activates the enzyme (Martin et al2000) This reaction leads to the production of cGMPandbrings aboutthe activation of cGMP-dependent kinases which transduce multiplesignaling events through protein phosphorylation (Murad 2006)Non-canonicalNO signaling is achievedmainly by S-nitrosylation Inthis reaction NO covalently binds to alkyl sulfur atoms on proteinsand organic compounds without the assistance of enzymes to formS-nitrosothiols This reaction requires higher concentrations of NOand tends to proceed with slower kinetics than cGMP-mediatedactions S-nitrosylation impacts protein function stability andlocalization by modulating the cysteine-containing active sites ofenzymes and by regulating protein-protein interactions throughaltering the affinity of cysteine-containing binding niches (Hesset al 2005 Gould et al 2013 Doulias et al 2013)NO can be further metabolized to form reactive nitrogen
species such as peroxynitrite (OONOminus) which have distinctivephysiological and pathological roles of their own (Adams et al2015) Peroxynitrite forms when NO reacts with superoxide itis released from immune cells to assist with pathogen killingvia the oxidization of protein residues However the overproductionor dysregulation of peroxynitrite levels can lead to a chronicinflammatory response (Adams et al 2015) In additionNOS generates superoxide and hydrogen peroxide when theconcentrations of arginine and BH4 are low (Porasuphatana et al2003) For example when vascular BH4 levels are limited electronflow to molecular oxygen becomes lsquouncoupledrsquo from arginineoxidation resulting in the generation of superoxide anion and ofother reactive oxygen species (ROS) rather than in the generation
of NO Both superoxide anions and ROS contribute to thepathogenesis of vascular disease (Chuaiphichai et al 2017) andto cancer progression as will be further discussed below (Rabenderet al 2015)
Thus NO-dependent signaling pathways are highly complex andthe proper regulation of NO production is vital for executing thefunctions of these pathways Central to this regulation is theproduction and availability of the NO precursor arginine
NO regulation by arginine metabolismAlthough the physiological intracellular concentrations of argininefar exceed those required for eNOS to synthesize NO the acuteprovision of exogenous arginine increases NO production(Dioguardi 2011) This phenomenon is known as the lsquoarginineparadoxrsquo The arginine pool derives from several sources includingdietary intake body protein breakdown and endogenous de novosynthesis (Fig 1) Endogenous systemic arginine production occursthrough the intestinal-renal axis in which citrulline produced by theintestine is converted into arginine by the kidneys Here citrullineavailability is the limiting factor for the amount of argininesynthesized (Wu et al 2009) To supply tissues with their requiredarginine needs circulating arginine traverses cell membranes viathe Na+-independent cationic amino acid transport system y+which regulates the availability of arginine for arginine-dependentsynthetic pathways (Arancibia-Garavilla et al 2003) Of notearginine is also produced as an intermediate in the liver urea cyclebut here it participates in the detoxification of excess nitrogen assuch it is not secreted into the plasma and does not contribute to thebodyrsquos pool of arginine (Watford 1991 Wu et al 2009)
Arginine is a semi-essential amino acid meaning that underphysiological conditions its endogenous synthesis is sufficient tomeet the body requirements and no additional supplementation isrequired from diet In certain physiological and pathological statessuch as during infancy growth pregnancy and illness such asinfections and cancer arginine is synthesized endogenously inmultiple tissues by the arginine-citrulline cycle This is because theamount of circulating arginine as derived from dietary intake andkidney production cannot meet cellular requirements for arginineduring these states (Fig 1) The arginine-citrulline cycle operates inmost mammalian cell types where arginine is generated to meetcellular needs for its downstream metabolites (Husson et al 2003)Indeed arginine is a major metabolic nexus for the synthesis ofmultiple metabolites among which are NO polyamines prolineand creatine all of which are essential for cell survival andproliferation (Rhee et al 2007 Liang et al 2013 Sestili et al2016) Besides NOS the other two enzymes that function in thearginine-citrulline cycle are argininosuccinate synthase 1 (ASS1)and argininosuccinate lyase (ASL) both of which also function inrenal arginine production and in the liver as part of the urea cycleASS1 is a cytosolic enzyme that catalyzes the formation ofargininosuccinate from citrulline and aspartate with ATP beingbroken down into AMP and pyrophosphate during the reactionSubsequently ASL promotes the cleavage of argininosuccinate toarginine and fumarate Arginine can then be recycled back tocitrulline by NOS or be utilized by other enzymes for the synthesisof either ornithine agmatine or guanidinoacetate In contrast to thefour cellular enzymes that use arginine as a substrate only ASL cangenerate endogenous arginine in mammalian cells (Fig 1) Thusanother important factor that regulates the intracellular availabilityof arginine for NOS is the activity of other competing enzymesthat also use arginine as a substrate One of the main competitorsis another urea cycle enzyme arginase which converts arginine
Box 1 GlossaryAnchorage independence the capacity of cancer cells to divide andfunction despite the absence of a stable surface to anchor toEpithelial-to-mesenchymal transition (EMT) a process by whichepithelial cells lose their cell polarity and cell-cell adhesion and gainmigratory and invasive properties to become mesenchymal-like cellsNO donor amolecule that induces nitric oxide (NO) under physiologicalconditionsS-nitrosylation a protein post-translational modification in which NO iscovalently attached to cysteine residues to form S-nitrosocysteineTetrahydrobiopterin (BH4) a naturally occurring cofactor that isrequired by nitric oxide synthase (NOS) enzymes to produce NO aswell as by other enzymes involved in amino acid degradation and inneurotransmitter synthesis
into ornithine and urea thereby limiting the availability of argininefor NOS (Caldwell et al 2015) There are two isozymes of thisenzyme arginase-1 (ARG1) which functions in the urea cycle andis located primarily in hepatocytes and arginase-2 (ARG2) whichis ubiquitously expressed outside of the liver where it competeswith NOS for arginine (Caldwell et al 2015) Importantlyin response to an inflammatory stimulus like LPS activated(M1) macrophages express iNOS whereas during inflammationresolution macrophages switch to express ARG1 which sequestersarginine from iNOS This switch in gene expression leads to theincreased production of ornithine and its downstream metabolitespolyamines and proline with a subsequent decrease in NOproduction (Weisser et al 2013)Thus various factors determine the availability of intracellular
arginine in the specific cellular compartment for the synthesis ofNO including dietary intake of arginine the expression of itstransporters its level of synthesis by ASS1 and ASL and itscompeting use as a substrate by other enzymes These multiplevariables impact NO levels during homeostasis as well as indifferent disease states such as cancer (Wu et al 2009)
Dichotomous roles of NO in cancerNO has been linked to the pathogenesis of different tumor typesfunctioning as either an enhancer or inhibitor of cancer development
(Fig 2) The contribution of NO to cancer progression includes theactivation of mitogenic pathways Treating cultured human breastcancer cells with an NO donor (Box 1) resulted in the activationof the epidermal growth factor receptor (EGFR) and of theextracellular signal-regulated kinase (ERK) pathways throughEGFR S-nitrosylation with a subsequent increase in the migrationand invasive potential of these cells (Garrido et al 2017) Similarlythe mTOR mitogenic pathway has been shown to be activated byNO via the S-nitrosylation of key proteins to promote theproliferation of human melanoma cells both in vitro and in ananimal xenograft models (Lopez-Rivera et al 2014) Anotherimportant oncogenic pathway in cancer promoted by NO to induceproliferation and migration is the Wntβ-catenin pathway assuggested by the activation of Wnt target genes following theoverexpression of iNOS in cultured human colon and breast cancercells (Du et al 2013) Interestingly NO can also support tumor-forming cancer stem cells (CSCs) as NO produced in culturedhuman colon CSCs was found to drive stemness-related signalingpathways central to colon tumor initiation and progression (Puglisiet al 2015) In contrast to the role that NO plays in supportingoncogenic pathways in cancer NO also exhibits an anti-proliferativerole by suppressing oncogenic pathways or by activating tumor-suppressing ones Indeed NO has been shown to negativelyregulate the proliferation of human neuroblastoma cell lines by
Dietary arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
Glutamate
Proline
PolyaminesUrea
AgmatineGuanidinoacetate
Creatine
iNOS
eNOS
nNOS
ASS1
NO
y+ transport s
ystem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
ADC
GAM
T
Arginase 2
Fig 1 A schematic illustration of argininemetabolism outside of the liverArginine from dietary intake can enter a cell via the y+ transport system or it can besynthesized endogenously by the arginine-citrulline cycle (red arrows) In contrast to the single enzyme that synthesizes arginine (ASL) many enzymesutilize arginine as their substrate (black arrows) to synthesize a range of compounds including ornithine agmatine guanidinoacetate andNO in accordancewithcellular needs NO is synthesized from arginine by either one or by all three NOS isoforms (eNOS iNOS or nNOS) depending on cellular context ADCarginine decarboxylase ASA argininosuccinic acid ASL argininosuccinate lyase ASS argininosuccinate synthase 1 eNOS endothelial nitric oxide synthaseGAMT guanidinoacetate methyltransferase iNOS inducible nitric oxide synthase NO nitric oxide nNOS neuronal nitric oxide synthase
decreasing the expression of the oncogene c-Myc in a cGMP-dependent manner (Ciani et al 2004) Moreover NO can inhibitthe proliferation of a human neuroblastoma cancer cell line in vitroby upregulating tumor suppressor pathways including the BRCA1Chk1p53 pathway leading to cell cycle arrest in response to DNAdamage via activation of cell cycle checkpoints (Van de Wouweret al 2012) Of note the dual roles of NO in cancer are also dose-dependent while exogenous NO stimulated cell proliferation inpheochromocytoma PC12 cells at low concentrations it inhibitedproliferation at higher concentrations (Bal-Price et al 2006)Likewise whereas low levels of NO can inhibit apoptosis andpromote cancer high levels of NO can contribute to cancer cellapoptosis (Villalobo 2006)NO has also been implicated in the epigenetic modification
of gene expression Several studies have described NO-drivenepigenetic modifications that control normal biologicaldevelopment and mediate tumorigenesis (Vasudevan et al 2016)In prostate carcinogenesis the silencing of glutathione transferaseP1-1 (GSTP1) is a common early event that is frequently caused bypromoter hypermethylation and correlates with decreased survivalIn human prostate cancer cells eNOS participates in GSTP1repression by being recruited to the gene promoter with aconsequential remodeling of the local chromatin (Re et al 2011)Importantly pharmacological inhibition of eNOS relieved therepression of GSTP1 and treatment with an NO donor silenced thisgene suggesting that eNOS regulates GSTP1 transcription throughNO production In oral squamous cell carcinoma patients it iscommon to find histone hyperacetylation that promotes tumorprogression Interestingly it has been found that NO mediateshistone hyperacetylation and that p300 histone acetylase activity isdependent on endogenously generated NO (Arif et al 2010)Moreover NO can affect histone PTMs at a global level astreatment with NO donors resulted in the differential expression of
over 6500 genes in breast cancer cells in which the pattern of PTMscorrelated with an oncogenic signature (Vasudevan et al 2015)Conversely NO has been found to inhibit lysine demethylase 3A(KDM3A) a histone demethylase that is known to positivelyregulate cancer cell invasion chemoresistance and metastasis inbreast and ovarian cancer cells (Hickok et al 2013)
The tumor microenvironment also influences the contribution ofNO to tumor fate Indeed cancer cells have developed diverse waysto intervene in the production andor metabolism of NO in tumorsand their surrounding tissue to gain an advantage As with itsdichotomous roles in cancer cell proliferation and apoptosis NOcan influence both cancer progression and its restriction NOS1upregulation was found to support the growth and activity ofcultured cancer-associated fibroblasts which are known to stimulatetumor progression (Augsten et al 2014) Additionally NO wasdemonstrated to directly promote cancer progression and invasionby influencing the stromal components of a tumor by inducingepithelial-to-mesenchymal transition (EMT Box 1) as well as byaffecting tumor vessel formation In human squamous cellcarcinoma and in lung cancer cells lines EMT and stem cellfeatures are reportedly activated by moderate levels of NO producedby NOS in response to growth factors and inflammatory mediators(Terzuoli et al 2017) On the other hand it seems that in the samecells higher than normal concentrations of NO inhibit EMT Inhuman metastatic prostate cancer cell lines treated with a highconcentration of the NO donor DETA-NONOate EMT and theinvasive phenotype of these cells is reversed via the inhibition of theEMT effector and transcription factor Snail (Baritaki et al 2010) Itwas demonstrated in this study that both Snail mRNA levels and itsDNA-binding capacity were inhibited by NO in a yet-undefinedmechanism Moreover NO was found to inhibit the mitochondrialfunction of Complex I and IV of the electron transport chain (Daiet al 2013) and to perturb the integrity of the mitochondrial
Growth factor induction
Oxidative DNA damage
Angiogenesis induction
Apoptosis inhibition
EMT induction
EMT inhibition
Apoptosis induction
Immune cell activation
Angiogenesis inhibition
Tumor suppressor pathway
activation
Cancer-promoting effects
Cancer-restrictingeffects
NO
Fig 2 Dichotomous roles for NO in cancer The dichotomousroles of NO in cancer are depicted in a Yin and Yang model toillustrate its cancer-promoting and cancer-restricting effectsNO can either inhibit or enhance cancer progression dependingon the biological context its concentration and on the duration ofNO production EMT epithelial-to-mesenchymal transitionNO nitric oxide
network by S-nitrosylation of dynamine-1-like protein (DRP-1)which is known to regulate mitochondrial fission (Cho et al 2009)It is thus tempting to speculate that the proliferation and EMTelicited by low doses of NO in cancer are associated with theinhibition of mitochondrial activity (Boland et al 2013)Cancer progression also depends on angiogenesis which is
needed to support the growing tumor with oxygen and nutrients andto remove waste products NO can promote angiogenesis bysupporting endothelial differentiation by inhibiting antiangiogenicfactors by dilating tumor blood vessels and by recruiting bone-marrow-derived and perivascular cells (Fukumura et al 2006) In amurine melanoma model lacking eNOS expression a perturbedrecruitment of mural cells to newly formed vessels and abnormalvessel branching and stabilization were demonstrated (Kashiwagiet al 2005) Moreover nNOS was required for the formationof abnormal tumor blood vessels in mice harboring humanglioma xenografts (Kashiwagi et al 2008) Conversely severalNO-related metabolites such as isosorbide mononitrate anddinitrate have been found to suppress VEGF protein levels incultured human colon cancer cells and to inhibit angiogenesisin vivo in xenografts of murine lung tumors (Pathi et al 2011Pipili-Synetos et al 1995)In recent years researchers have focused on the role of the
immune system in cancer development Here too NO produced byimmune cells has dual regulatory functions in tumor progressionNO and reactive nitrogen species that originate from immune cellssuch as macrophages and neutrophils can have pro-tumorigeniceffects on neighboring epithelial cells for example by inducingDNA damage that can initiate inflammation-associated neoplastictransformation (Wang et al 2017) NO produced by tumor-infiltrating myeloid cells was found to be important for activation ofadoptively transferred cytotoxic T cells (Marigo et al 2016) It wasalso shown that NO production by colon cells is required inpathogen-induced colon inflammation and immune cell infiltrationeventually leading to dysplasia and colon cancer development(Erdman et al 2009) In parallel NO can activate macrophages andcytotoxic T cells and augment the immune response against tumorcells (MacMicking et al 1997 Marigo et al 2016) Indeed thecytotoxic action of the cytokine interferon gamma (IFN-γ) or theactivity of LPS-activated primary mouse macrophages againstdifferent cancer cell lines were impaired in macrophages fromNOS2 knock-out mice (MacMicking et al 1997) Moreover animalstudies have demonstrated that in tumor vessels of melanomaxenografts macrophage-derived NO induced the expression of theadhesion molecule VCAM-1 which is important for T-cellextravasation Additionally only co-transfer of CD8+ T cells withwild-type macrophages but not withNos2minusminusmacrophages yieldedT-cell homing to the tumor and consequently led to tumor rejection(Sektioglu et al 2016)These studies collectively suggest that cellular levels of NO and
the associated cross-talk between cancer cells and theirenvironment are important for tumor initiation and progressionAs a consequence efforts have beenmade in recent years to improveNO detection and analysis (Box 2)
Metabolic regulation of NO synthesis in cancerCancer cells regulate the availability of intracellular arginine for NOsynthesis in multiple ways Cultured human colon cancer cells andmurine breast cancer cells stimulated by inflammatory mediatorssuch as LPS and IFN-γ increase the availability of arginine for NOproduction by enhancing transmembrane arginine import (Cendanet al 1996ab) Omental adipose stromal cells (O-ASCs) are
mesenchymal stem cells contained in the omentum tissue that areknown to promote endometrial and ovarian tumor proliferationInterestingly O-ASCs can support human endometrial or ovariancancer cells in co-culture by supplying them with arginine for NOproduction a finding that may reflect their role in tumor progression(Salimian Rizi et al 2015) Another way to upregulate argininelevels in tumor cells is to increase its endogenous synthesis Insamples from colon and breast cancer patients the overexpressionof ASL which encodes the enzyme that synthesizes arginineis associated with poor survival (Huang et al 2015 2017a)Additionally tumor samples from hepatocellular carcinoma patientsshow increased ASL expression and in human colon breast andhepatocellular carcinoma cells silencing of ASL expression byshort-hairpin RNA or the reduction of NO production by a NOSinhibitor inhibited cancer cellsrsquo proliferation and anchorage-independence (Box 1) (Huang et al 2013 2015 2017a) Thesestudies imply that ASL exerts its tumorigenic effects at least in partthrough NO but they do not rule out the possibility that increasedarginine levels might promote cancer by supporting the synthesis ofother arginine-derived molecules such as polyamines
Another urea cycle enzyme that functions in the arginine-citrulline cycle ASS1 is also overexpressed in various humancancers including in lung colon gastric and ovarian cancer(Delage et al 2010) The upregulated expression of ASS1 has beenfound to support the proliferation of human colon cancer cellsin vitro as well as the migration and metastatic potential of humangastric cell lines both in vitro and in mice with tumor xenografts(Bateman et al 2017 Shan et al 2015) Yet the cancer-promotingmechanisms fostered by ASS1 overexpression and their clinicalimplications remain unclear It is possible that high levels of ASS1support tumor proliferation and aggressiveness by increasing thesupply of arginine for NO production In support of this ASS1overexpression in rat vascular smooth muscle cells potentiated theLPS- and IFN-γ-stimulated production of NO (Xie and Gross1997) Moreover human breast cancer cells treated with the pro-inflammatory cytokine interleukin 17 (IL-17) increased theirproliferation and this response was reported to depend on the
Box 2 Detecting NO in cancerCellular concentrations of nitric oxide (NO) are hard to quantify due to itsvery short half-life In biological tissue samples NO levels can bemeasured indirectly by quantifying nitrite and nitrate using high-performance liquid chromatography (HPLC) (Jiang et al 2012) Thereagent 4-amino-5-methylamino-2prime7prime-difluorofluorescein (DAF-FM) isused for intracellular measurements because it becomes fluorescentwhen it reacts with NO and can be detected by any fluorescein-detectinginstrument (Namin et al 2013) Undoubtedly the ideal method formeasuring NO levels is via a flux analysis performed in vivo by usinglabeled arginine and then tracing the downstream products such ascitrulline and NO-derived urinary nitrate by using gas or liquidchromatography-mass spectrometry (Magneacute et al 2009)For a more global assessment of NO involvement in cancer the
expression of individual genes in the arginine-NO pathway can beassessed across different types of cancer in large datasets such as inThe Cancer Genome Atlas (TCGA) Expression levels can then becorrelated with patient survival and therapeutic response (Ekmekciogluet al 2016)Although these methods have all contributed to our understanding of
NO biology NO production and its downstream effects often involvecomplex signaling pathways and thus it remains challenging to dissectthe exact cellular contribution of NO to disease pathogenesis and toinvestigate its therapeutic relevance
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
(Lundberg et al 2008) and hence will not be discussed furtherhereCellular NO levels are tightly regulated at several different levels
and by multiple factors NO can be regulated at the level of NOStranscription or via the post-translational modification of NOS aswell as via the cellular expression of the different NOS isoformsand through the availability of NOS substrates such as arginine andBH4 (Mungrue et al 2003) It can also be regulated by the amountof NO produced by the different NOS as well as by the short half-life of NO which is estimated to be in the range of 01ndash2 s andallows the rapid termination of NO signaling cascades once theinitial stimulus is turned off (Thomas et al 2001)The biological effects of NO are exerted through either cyclic
guanosine monophosphate (cGMP) or via post-translationalmodification (PTM) by S-nitrosylation (Box 1) (Seth et al 2018)Canonical NO signaling involves soluble guanylate cyclase (sGC)which is the only known receptor for NO This enzyme is aheterodimer composed of two subunits one ofwhich contains a hemegroup to which NO binds and activates the enzyme (Martin et al2000) This reaction leads to the production of cGMPandbrings aboutthe activation of cGMP-dependent kinases which transduce multiplesignaling events through protein phosphorylation (Murad 2006)Non-canonicalNO signaling is achievedmainly by S-nitrosylation Inthis reaction NO covalently binds to alkyl sulfur atoms on proteinsand organic compounds without the assistance of enzymes to formS-nitrosothiols This reaction requires higher concentrations of NOand tends to proceed with slower kinetics than cGMP-mediatedactions S-nitrosylation impacts protein function stability andlocalization by modulating the cysteine-containing active sites ofenzymes and by regulating protein-protein interactions throughaltering the affinity of cysteine-containing binding niches (Hesset al 2005 Gould et al 2013 Doulias et al 2013)NO can be further metabolized to form reactive nitrogen
species such as peroxynitrite (OONOminus) which have distinctivephysiological and pathological roles of their own (Adams et al2015) Peroxynitrite forms when NO reacts with superoxide itis released from immune cells to assist with pathogen killingvia the oxidization of protein residues However the overproductionor dysregulation of peroxynitrite levels can lead to a chronicinflammatory response (Adams et al 2015) In additionNOS generates superoxide and hydrogen peroxide when theconcentrations of arginine and BH4 are low (Porasuphatana et al2003) For example when vascular BH4 levels are limited electronflow to molecular oxygen becomes lsquouncoupledrsquo from arginineoxidation resulting in the generation of superoxide anion and ofother reactive oxygen species (ROS) rather than in the generation
of NO Both superoxide anions and ROS contribute to thepathogenesis of vascular disease (Chuaiphichai et al 2017) andto cancer progression as will be further discussed below (Rabenderet al 2015)
Thus NO-dependent signaling pathways are highly complex andthe proper regulation of NO production is vital for executing thefunctions of these pathways Central to this regulation is theproduction and availability of the NO precursor arginine
NO regulation by arginine metabolismAlthough the physiological intracellular concentrations of argininefar exceed those required for eNOS to synthesize NO the acuteprovision of exogenous arginine increases NO production(Dioguardi 2011) This phenomenon is known as the lsquoarginineparadoxrsquo The arginine pool derives from several sources includingdietary intake body protein breakdown and endogenous de novosynthesis (Fig 1) Endogenous systemic arginine production occursthrough the intestinal-renal axis in which citrulline produced by theintestine is converted into arginine by the kidneys Here citrullineavailability is the limiting factor for the amount of argininesynthesized (Wu et al 2009) To supply tissues with their requiredarginine needs circulating arginine traverses cell membranes viathe Na+-independent cationic amino acid transport system y+which regulates the availability of arginine for arginine-dependentsynthetic pathways (Arancibia-Garavilla et al 2003) Of notearginine is also produced as an intermediate in the liver urea cyclebut here it participates in the detoxification of excess nitrogen assuch it is not secreted into the plasma and does not contribute to thebodyrsquos pool of arginine (Watford 1991 Wu et al 2009)
Arginine is a semi-essential amino acid meaning that underphysiological conditions its endogenous synthesis is sufficient tomeet the body requirements and no additional supplementation isrequired from diet In certain physiological and pathological statessuch as during infancy growth pregnancy and illness such asinfections and cancer arginine is synthesized endogenously inmultiple tissues by the arginine-citrulline cycle This is because theamount of circulating arginine as derived from dietary intake andkidney production cannot meet cellular requirements for arginineduring these states (Fig 1) The arginine-citrulline cycle operates inmost mammalian cell types where arginine is generated to meetcellular needs for its downstream metabolites (Husson et al 2003)Indeed arginine is a major metabolic nexus for the synthesis ofmultiple metabolites among which are NO polyamines prolineand creatine all of which are essential for cell survival andproliferation (Rhee et al 2007 Liang et al 2013 Sestili et al2016) Besides NOS the other two enzymes that function in thearginine-citrulline cycle are argininosuccinate synthase 1 (ASS1)and argininosuccinate lyase (ASL) both of which also function inrenal arginine production and in the liver as part of the urea cycleASS1 is a cytosolic enzyme that catalyzes the formation ofargininosuccinate from citrulline and aspartate with ATP beingbroken down into AMP and pyrophosphate during the reactionSubsequently ASL promotes the cleavage of argininosuccinate toarginine and fumarate Arginine can then be recycled back tocitrulline by NOS or be utilized by other enzymes for the synthesisof either ornithine agmatine or guanidinoacetate In contrast to thefour cellular enzymes that use arginine as a substrate only ASL cangenerate endogenous arginine in mammalian cells (Fig 1) Thusanother important factor that regulates the intracellular availabilityof arginine for NOS is the activity of other competing enzymesthat also use arginine as a substrate One of the main competitorsis another urea cycle enzyme arginase which converts arginine
Box 1 GlossaryAnchorage independence the capacity of cancer cells to divide andfunction despite the absence of a stable surface to anchor toEpithelial-to-mesenchymal transition (EMT) a process by whichepithelial cells lose their cell polarity and cell-cell adhesion and gainmigratory and invasive properties to become mesenchymal-like cellsNO donor amolecule that induces nitric oxide (NO) under physiologicalconditionsS-nitrosylation a protein post-translational modification in which NO iscovalently attached to cysteine residues to form S-nitrosocysteineTetrahydrobiopterin (BH4) a naturally occurring cofactor that isrequired by nitric oxide synthase (NOS) enzymes to produce NO aswell as by other enzymes involved in amino acid degradation and inneurotransmitter synthesis
into ornithine and urea thereby limiting the availability of argininefor NOS (Caldwell et al 2015) There are two isozymes of thisenzyme arginase-1 (ARG1) which functions in the urea cycle andis located primarily in hepatocytes and arginase-2 (ARG2) whichis ubiquitously expressed outside of the liver where it competeswith NOS for arginine (Caldwell et al 2015) Importantlyin response to an inflammatory stimulus like LPS activated(M1) macrophages express iNOS whereas during inflammationresolution macrophages switch to express ARG1 which sequestersarginine from iNOS This switch in gene expression leads to theincreased production of ornithine and its downstream metabolitespolyamines and proline with a subsequent decrease in NOproduction (Weisser et al 2013)Thus various factors determine the availability of intracellular
arginine in the specific cellular compartment for the synthesis ofNO including dietary intake of arginine the expression of itstransporters its level of synthesis by ASS1 and ASL and itscompeting use as a substrate by other enzymes These multiplevariables impact NO levels during homeostasis as well as indifferent disease states such as cancer (Wu et al 2009)
Dichotomous roles of NO in cancerNO has been linked to the pathogenesis of different tumor typesfunctioning as either an enhancer or inhibitor of cancer development
(Fig 2) The contribution of NO to cancer progression includes theactivation of mitogenic pathways Treating cultured human breastcancer cells with an NO donor (Box 1) resulted in the activationof the epidermal growth factor receptor (EGFR) and of theextracellular signal-regulated kinase (ERK) pathways throughEGFR S-nitrosylation with a subsequent increase in the migrationand invasive potential of these cells (Garrido et al 2017) Similarlythe mTOR mitogenic pathway has been shown to be activated byNO via the S-nitrosylation of key proteins to promote theproliferation of human melanoma cells both in vitro and in ananimal xenograft models (Lopez-Rivera et al 2014) Anotherimportant oncogenic pathway in cancer promoted by NO to induceproliferation and migration is the Wntβ-catenin pathway assuggested by the activation of Wnt target genes following theoverexpression of iNOS in cultured human colon and breast cancercells (Du et al 2013) Interestingly NO can also support tumor-forming cancer stem cells (CSCs) as NO produced in culturedhuman colon CSCs was found to drive stemness-related signalingpathways central to colon tumor initiation and progression (Puglisiet al 2015) In contrast to the role that NO plays in supportingoncogenic pathways in cancer NO also exhibits an anti-proliferativerole by suppressing oncogenic pathways or by activating tumor-suppressing ones Indeed NO has been shown to negativelyregulate the proliferation of human neuroblastoma cell lines by
Dietary arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
Glutamate
Proline
PolyaminesUrea
AgmatineGuanidinoacetate
Creatine
iNOS
eNOS
nNOS
ASS1
NO
y+ transport s
ystem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
ADC
GAM
T
Arginase 2
Fig 1 A schematic illustration of argininemetabolism outside of the liverArginine from dietary intake can enter a cell via the y+ transport system or it can besynthesized endogenously by the arginine-citrulline cycle (red arrows) In contrast to the single enzyme that synthesizes arginine (ASL) many enzymesutilize arginine as their substrate (black arrows) to synthesize a range of compounds including ornithine agmatine guanidinoacetate andNO in accordancewithcellular needs NO is synthesized from arginine by either one or by all three NOS isoforms (eNOS iNOS or nNOS) depending on cellular context ADCarginine decarboxylase ASA argininosuccinic acid ASL argininosuccinate lyase ASS argininosuccinate synthase 1 eNOS endothelial nitric oxide synthaseGAMT guanidinoacetate methyltransferase iNOS inducible nitric oxide synthase NO nitric oxide nNOS neuronal nitric oxide synthase
decreasing the expression of the oncogene c-Myc in a cGMP-dependent manner (Ciani et al 2004) Moreover NO can inhibitthe proliferation of a human neuroblastoma cancer cell line in vitroby upregulating tumor suppressor pathways including the BRCA1Chk1p53 pathway leading to cell cycle arrest in response to DNAdamage via activation of cell cycle checkpoints (Van de Wouweret al 2012) Of note the dual roles of NO in cancer are also dose-dependent while exogenous NO stimulated cell proliferation inpheochromocytoma PC12 cells at low concentrations it inhibitedproliferation at higher concentrations (Bal-Price et al 2006)Likewise whereas low levels of NO can inhibit apoptosis andpromote cancer high levels of NO can contribute to cancer cellapoptosis (Villalobo 2006)NO has also been implicated in the epigenetic modification
of gene expression Several studies have described NO-drivenepigenetic modifications that control normal biologicaldevelopment and mediate tumorigenesis (Vasudevan et al 2016)In prostate carcinogenesis the silencing of glutathione transferaseP1-1 (GSTP1) is a common early event that is frequently caused bypromoter hypermethylation and correlates with decreased survivalIn human prostate cancer cells eNOS participates in GSTP1repression by being recruited to the gene promoter with aconsequential remodeling of the local chromatin (Re et al 2011)Importantly pharmacological inhibition of eNOS relieved therepression of GSTP1 and treatment with an NO donor silenced thisgene suggesting that eNOS regulates GSTP1 transcription throughNO production In oral squamous cell carcinoma patients it iscommon to find histone hyperacetylation that promotes tumorprogression Interestingly it has been found that NO mediateshistone hyperacetylation and that p300 histone acetylase activity isdependent on endogenously generated NO (Arif et al 2010)Moreover NO can affect histone PTMs at a global level astreatment with NO donors resulted in the differential expression of
over 6500 genes in breast cancer cells in which the pattern of PTMscorrelated with an oncogenic signature (Vasudevan et al 2015)Conversely NO has been found to inhibit lysine demethylase 3A(KDM3A) a histone demethylase that is known to positivelyregulate cancer cell invasion chemoresistance and metastasis inbreast and ovarian cancer cells (Hickok et al 2013)
The tumor microenvironment also influences the contribution ofNO to tumor fate Indeed cancer cells have developed diverse waysto intervene in the production andor metabolism of NO in tumorsand their surrounding tissue to gain an advantage As with itsdichotomous roles in cancer cell proliferation and apoptosis NOcan influence both cancer progression and its restriction NOS1upregulation was found to support the growth and activity ofcultured cancer-associated fibroblasts which are known to stimulatetumor progression (Augsten et al 2014) Additionally NO wasdemonstrated to directly promote cancer progression and invasionby influencing the stromal components of a tumor by inducingepithelial-to-mesenchymal transition (EMT Box 1) as well as byaffecting tumor vessel formation In human squamous cellcarcinoma and in lung cancer cells lines EMT and stem cellfeatures are reportedly activated by moderate levels of NO producedby NOS in response to growth factors and inflammatory mediators(Terzuoli et al 2017) On the other hand it seems that in the samecells higher than normal concentrations of NO inhibit EMT Inhuman metastatic prostate cancer cell lines treated with a highconcentration of the NO donor DETA-NONOate EMT and theinvasive phenotype of these cells is reversed via the inhibition of theEMT effector and transcription factor Snail (Baritaki et al 2010) Itwas demonstrated in this study that both Snail mRNA levels and itsDNA-binding capacity were inhibited by NO in a yet-undefinedmechanism Moreover NO was found to inhibit the mitochondrialfunction of Complex I and IV of the electron transport chain (Daiet al 2013) and to perturb the integrity of the mitochondrial
Growth factor induction
Oxidative DNA damage
Angiogenesis induction
Apoptosis inhibition
EMT induction
EMT inhibition
Apoptosis induction
Immune cell activation
Angiogenesis inhibition
Tumor suppressor pathway
activation
Cancer-promoting effects
Cancer-restrictingeffects
NO
Fig 2 Dichotomous roles for NO in cancer The dichotomousroles of NO in cancer are depicted in a Yin and Yang model toillustrate its cancer-promoting and cancer-restricting effectsNO can either inhibit or enhance cancer progression dependingon the biological context its concentration and on the duration ofNO production EMT epithelial-to-mesenchymal transitionNO nitric oxide
network by S-nitrosylation of dynamine-1-like protein (DRP-1)which is known to regulate mitochondrial fission (Cho et al 2009)It is thus tempting to speculate that the proliferation and EMTelicited by low doses of NO in cancer are associated with theinhibition of mitochondrial activity (Boland et al 2013)Cancer progression also depends on angiogenesis which is
needed to support the growing tumor with oxygen and nutrients andto remove waste products NO can promote angiogenesis bysupporting endothelial differentiation by inhibiting antiangiogenicfactors by dilating tumor blood vessels and by recruiting bone-marrow-derived and perivascular cells (Fukumura et al 2006) In amurine melanoma model lacking eNOS expression a perturbedrecruitment of mural cells to newly formed vessels and abnormalvessel branching and stabilization were demonstrated (Kashiwagiet al 2005) Moreover nNOS was required for the formationof abnormal tumor blood vessels in mice harboring humanglioma xenografts (Kashiwagi et al 2008) Conversely severalNO-related metabolites such as isosorbide mononitrate anddinitrate have been found to suppress VEGF protein levels incultured human colon cancer cells and to inhibit angiogenesisin vivo in xenografts of murine lung tumors (Pathi et al 2011Pipili-Synetos et al 1995)In recent years researchers have focused on the role of the
immune system in cancer development Here too NO produced byimmune cells has dual regulatory functions in tumor progressionNO and reactive nitrogen species that originate from immune cellssuch as macrophages and neutrophils can have pro-tumorigeniceffects on neighboring epithelial cells for example by inducingDNA damage that can initiate inflammation-associated neoplastictransformation (Wang et al 2017) NO produced by tumor-infiltrating myeloid cells was found to be important for activation ofadoptively transferred cytotoxic T cells (Marigo et al 2016) It wasalso shown that NO production by colon cells is required inpathogen-induced colon inflammation and immune cell infiltrationeventually leading to dysplasia and colon cancer development(Erdman et al 2009) In parallel NO can activate macrophages andcytotoxic T cells and augment the immune response against tumorcells (MacMicking et al 1997 Marigo et al 2016) Indeed thecytotoxic action of the cytokine interferon gamma (IFN-γ) or theactivity of LPS-activated primary mouse macrophages againstdifferent cancer cell lines were impaired in macrophages fromNOS2 knock-out mice (MacMicking et al 1997) Moreover animalstudies have demonstrated that in tumor vessels of melanomaxenografts macrophage-derived NO induced the expression of theadhesion molecule VCAM-1 which is important for T-cellextravasation Additionally only co-transfer of CD8+ T cells withwild-type macrophages but not withNos2minusminusmacrophages yieldedT-cell homing to the tumor and consequently led to tumor rejection(Sektioglu et al 2016)These studies collectively suggest that cellular levels of NO and
the associated cross-talk between cancer cells and theirenvironment are important for tumor initiation and progressionAs a consequence efforts have beenmade in recent years to improveNO detection and analysis (Box 2)
Metabolic regulation of NO synthesis in cancerCancer cells regulate the availability of intracellular arginine for NOsynthesis in multiple ways Cultured human colon cancer cells andmurine breast cancer cells stimulated by inflammatory mediatorssuch as LPS and IFN-γ increase the availability of arginine for NOproduction by enhancing transmembrane arginine import (Cendanet al 1996ab) Omental adipose stromal cells (O-ASCs) are
mesenchymal stem cells contained in the omentum tissue that areknown to promote endometrial and ovarian tumor proliferationInterestingly O-ASCs can support human endometrial or ovariancancer cells in co-culture by supplying them with arginine for NOproduction a finding that may reflect their role in tumor progression(Salimian Rizi et al 2015) Another way to upregulate argininelevels in tumor cells is to increase its endogenous synthesis Insamples from colon and breast cancer patients the overexpressionof ASL which encodes the enzyme that synthesizes arginineis associated with poor survival (Huang et al 2015 2017a)Additionally tumor samples from hepatocellular carcinoma patientsshow increased ASL expression and in human colon breast andhepatocellular carcinoma cells silencing of ASL expression byshort-hairpin RNA or the reduction of NO production by a NOSinhibitor inhibited cancer cellsrsquo proliferation and anchorage-independence (Box 1) (Huang et al 2013 2015 2017a) Thesestudies imply that ASL exerts its tumorigenic effects at least in partthrough NO but they do not rule out the possibility that increasedarginine levels might promote cancer by supporting the synthesis ofother arginine-derived molecules such as polyamines
Another urea cycle enzyme that functions in the arginine-citrulline cycle ASS1 is also overexpressed in various humancancers including in lung colon gastric and ovarian cancer(Delage et al 2010) The upregulated expression of ASS1 has beenfound to support the proliferation of human colon cancer cellsin vitro as well as the migration and metastatic potential of humangastric cell lines both in vitro and in mice with tumor xenografts(Bateman et al 2017 Shan et al 2015) Yet the cancer-promotingmechanisms fostered by ASS1 overexpression and their clinicalimplications remain unclear It is possible that high levels of ASS1support tumor proliferation and aggressiveness by increasing thesupply of arginine for NO production In support of this ASS1overexpression in rat vascular smooth muscle cells potentiated theLPS- and IFN-γ-stimulated production of NO (Xie and Gross1997) Moreover human breast cancer cells treated with the pro-inflammatory cytokine interleukin 17 (IL-17) increased theirproliferation and this response was reported to depend on the
Box 2 Detecting NO in cancerCellular concentrations of nitric oxide (NO) are hard to quantify due to itsvery short half-life In biological tissue samples NO levels can bemeasured indirectly by quantifying nitrite and nitrate using high-performance liquid chromatography (HPLC) (Jiang et al 2012) Thereagent 4-amino-5-methylamino-2prime7prime-difluorofluorescein (DAF-FM) isused for intracellular measurements because it becomes fluorescentwhen it reacts with NO and can be detected by any fluorescein-detectinginstrument (Namin et al 2013) Undoubtedly the ideal method formeasuring NO levels is via a flux analysis performed in vivo by usinglabeled arginine and then tracing the downstream products such ascitrulline and NO-derived urinary nitrate by using gas or liquidchromatography-mass spectrometry (Magneacute et al 2009)For a more global assessment of NO involvement in cancer the
expression of individual genes in the arginine-NO pathway can beassessed across different types of cancer in large datasets such as inThe Cancer Genome Atlas (TCGA) Expression levels can then becorrelated with patient survival and therapeutic response (Ekmekciogluet al 2016)Although these methods have all contributed to our understanding of
NO biology NO production and its downstream effects often involvecomplex signaling pathways and thus it remains challenging to dissectthe exact cellular contribution of NO to disease pathogenesis and toinvestigate its therapeutic relevance
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
into ornithine and urea thereby limiting the availability of argininefor NOS (Caldwell et al 2015) There are two isozymes of thisenzyme arginase-1 (ARG1) which functions in the urea cycle andis located primarily in hepatocytes and arginase-2 (ARG2) whichis ubiquitously expressed outside of the liver where it competeswith NOS for arginine (Caldwell et al 2015) Importantlyin response to an inflammatory stimulus like LPS activated(M1) macrophages express iNOS whereas during inflammationresolution macrophages switch to express ARG1 which sequestersarginine from iNOS This switch in gene expression leads to theincreased production of ornithine and its downstream metabolitespolyamines and proline with a subsequent decrease in NOproduction (Weisser et al 2013)Thus various factors determine the availability of intracellular
arginine in the specific cellular compartment for the synthesis ofNO including dietary intake of arginine the expression of itstransporters its level of synthesis by ASS1 and ASL and itscompeting use as a substrate by other enzymes These multiplevariables impact NO levels during homeostasis as well as indifferent disease states such as cancer (Wu et al 2009)
Dichotomous roles of NO in cancerNO has been linked to the pathogenesis of different tumor typesfunctioning as either an enhancer or inhibitor of cancer development
(Fig 2) The contribution of NO to cancer progression includes theactivation of mitogenic pathways Treating cultured human breastcancer cells with an NO donor (Box 1) resulted in the activationof the epidermal growth factor receptor (EGFR) and of theextracellular signal-regulated kinase (ERK) pathways throughEGFR S-nitrosylation with a subsequent increase in the migrationand invasive potential of these cells (Garrido et al 2017) Similarlythe mTOR mitogenic pathway has been shown to be activated byNO via the S-nitrosylation of key proteins to promote theproliferation of human melanoma cells both in vitro and in ananimal xenograft models (Lopez-Rivera et al 2014) Anotherimportant oncogenic pathway in cancer promoted by NO to induceproliferation and migration is the Wntβ-catenin pathway assuggested by the activation of Wnt target genes following theoverexpression of iNOS in cultured human colon and breast cancercells (Du et al 2013) Interestingly NO can also support tumor-forming cancer stem cells (CSCs) as NO produced in culturedhuman colon CSCs was found to drive stemness-related signalingpathways central to colon tumor initiation and progression (Puglisiet al 2015) In contrast to the role that NO plays in supportingoncogenic pathways in cancer NO also exhibits an anti-proliferativerole by suppressing oncogenic pathways or by activating tumor-suppressing ones Indeed NO has been shown to negativelyregulate the proliferation of human neuroblastoma cell lines by
Dietary arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
Glutamate
Proline
PolyaminesUrea
AgmatineGuanidinoacetate
Creatine
iNOS
eNOS
nNOS
ASS1
NO
y+ transport s
ystem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
ADC
GAM
T
Arginase 2
Fig 1 A schematic illustration of argininemetabolism outside of the liverArginine from dietary intake can enter a cell via the y+ transport system or it can besynthesized endogenously by the arginine-citrulline cycle (red arrows) In contrast to the single enzyme that synthesizes arginine (ASL) many enzymesutilize arginine as their substrate (black arrows) to synthesize a range of compounds including ornithine agmatine guanidinoacetate andNO in accordancewithcellular needs NO is synthesized from arginine by either one or by all three NOS isoforms (eNOS iNOS or nNOS) depending on cellular context ADCarginine decarboxylase ASA argininosuccinic acid ASL argininosuccinate lyase ASS argininosuccinate synthase 1 eNOS endothelial nitric oxide synthaseGAMT guanidinoacetate methyltransferase iNOS inducible nitric oxide synthase NO nitric oxide nNOS neuronal nitric oxide synthase
decreasing the expression of the oncogene c-Myc in a cGMP-dependent manner (Ciani et al 2004) Moreover NO can inhibitthe proliferation of a human neuroblastoma cancer cell line in vitroby upregulating tumor suppressor pathways including the BRCA1Chk1p53 pathway leading to cell cycle arrest in response to DNAdamage via activation of cell cycle checkpoints (Van de Wouweret al 2012) Of note the dual roles of NO in cancer are also dose-dependent while exogenous NO stimulated cell proliferation inpheochromocytoma PC12 cells at low concentrations it inhibitedproliferation at higher concentrations (Bal-Price et al 2006)Likewise whereas low levels of NO can inhibit apoptosis andpromote cancer high levels of NO can contribute to cancer cellapoptosis (Villalobo 2006)NO has also been implicated in the epigenetic modification
of gene expression Several studies have described NO-drivenepigenetic modifications that control normal biologicaldevelopment and mediate tumorigenesis (Vasudevan et al 2016)In prostate carcinogenesis the silencing of glutathione transferaseP1-1 (GSTP1) is a common early event that is frequently caused bypromoter hypermethylation and correlates with decreased survivalIn human prostate cancer cells eNOS participates in GSTP1repression by being recruited to the gene promoter with aconsequential remodeling of the local chromatin (Re et al 2011)Importantly pharmacological inhibition of eNOS relieved therepression of GSTP1 and treatment with an NO donor silenced thisgene suggesting that eNOS regulates GSTP1 transcription throughNO production In oral squamous cell carcinoma patients it iscommon to find histone hyperacetylation that promotes tumorprogression Interestingly it has been found that NO mediateshistone hyperacetylation and that p300 histone acetylase activity isdependent on endogenously generated NO (Arif et al 2010)Moreover NO can affect histone PTMs at a global level astreatment with NO donors resulted in the differential expression of
over 6500 genes in breast cancer cells in which the pattern of PTMscorrelated with an oncogenic signature (Vasudevan et al 2015)Conversely NO has been found to inhibit lysine demethylase 3A(KDM3A) a histone demethylase that is known to positivelyregulate cancer cell invasion chemoresistance and metastasis inbreast and ovarian cancer cells (Hickok et al 2013)
The tumor microenvironment also influences the contribution ofNO to tumor fate Indeed cancer cells have developed diverse waysto intervene in the production andor metabolism of NO in tumorsand their surrounding tissue to gain an advantage As with itsdichotomous roles in cancer cell proliferation and apoptosis NOcan influence both cancer progression and its restriction NOS1upregulation was found to support the growth and activity ofcultured cancer-associated fibroblasts which are known to stimulatetumor progression (Augsten et al 2014) Additionally NO wasdemonstrated to directly promote cancer progression and invasionby influencing the stromal components of a tumor by inducingepithelial-to-mesenchymal transition (EMT Box 1) as well as byaffecting tumor vessel formation In human squamous cellcarcinoma and in lung cancer cells lines EMT and stem cellfeatures are reportedly activated by moderate levels of NO producedby NOS in response to growth factors and inflammatory mediators(Terzuoli et al 2017) On the other hand it seems that in the samecells higher than normal concentrations of NO inhibit EMT Inhuman metastatic prostate cancer cell lines treated with a highconcentration of the NO donor DETA-NONOate EMT and theinvasive phenotype of these cells is reversed via the inhibition of theEMT effector and transcription factor Snail (Baritaki et al 2010) Itwas demonstrated in this study that both Snail mRNA levels and itsDNA-binding capacity were inhibited by NO in a yet-undefinedmechanism Moreover NO was found to inhibit the mitochondrialfunction of Complex I and IV of the electron transport chain (Daiet al 2013) and to perturb the integrity of the mitochondrial
Growth factor induction
Oxidative DNA damage
Angiogenesis induction
Apoptosis inhibition
EMT induction
EMT inhibition
Apoptosis induction
Immune cell activation
Angiogenesis inhibition
Tumor suppressor pathway
activation
Cancer-promoting effects
Cancer-restrictingeffects
NO
Fig 2 Dichotomous roles for NO in cancer The dichotomousroles of NO in cancer are depicted in a Yin and Yang model toillustrate its cancer-promoting and cancer-restricting effectsNO can either inhibit or enhance cancer progression dependingon the biological context its concentration and on the duration ofNO production EMT epithelial-to-mesenchymal transitionNO nitric oxide
network by S-nitrosylation of dynamine-1-like protein (DRP-1)which is known to regulate mitochondrial fission (Cho et al 2009)It is thus tempting to speculate that the proliferation and EMTelicited by low doses of NO in cancer are associated with theinhibition of mitochondrial activity (Boland et al 2013)Cancer progression also depends on angiogenesis which is
needed to support the growing tumor with oxygen and nutrients andto remove waste products NO can promote angiogenesis bysupporting endothelial differentiation by inhibiting antiangiogenicfactors by dilating tumor blood vessels and by recruiting bone-marrow-derived and perivascular cells (Fukumura et al 2006) In amurine melanoma model lacking eNOS expression a perturbedrecruitment of mural cells to newly formed vessels and abnormalvessel branching and stabilization were demonstrated (Kashiwagiet al 2005) Moreover nNOS was required for the formationof abnormal tumor blood vessels in mice harboring humanglioma xenografts (Kashiwagi et al 2008) Conversely severalNO-related metabolites such as isosorbide mononitrate anddinitrate have been found to suppress VEGF protein levels incultured human colon cancer cells and to inhibit angiogenesisin vivo in xenografts of murine lung tumors (Pathi et al 2011Pipili-Synetos et al 1995)In recent years researchers have focused on the role of the
immune system in cancer development Here too NO produced byimmune cells has dual regulatory functions in tumor progressionNO and reactive nitrogen species that originate from immune cellssuch as macrophages and neutrophils can have pro-tumorigeniceffects on neighboring epithelial cells for example by inducingDNA damage that can initiate inflammation-associated neoplastictransformation (Wang et al 2017) NO produced by tumor-infiltrating myeloid cells was found to be important for activation ofadoptively transferred cytotoxic T cells (Marigo et al 2016) It wasalso shown that NO production by colon cells is required inpathogen-induced colon inflammation and immune cell infiltrationeventually leading to dysplasia and colon cancer development(Erdman et al 2009) In parallel NO can activate macrophages andcytotoxic T cells and augment the immune response against tumorcells (MacMicking et al 1997 Marigo et al 2016) Indeed thecytotoxic action of the cytokine interferon gamma (IFN-γ) or theactivity of LPS-activated primary mouse macrophages againstdifferent cancer cell lines were impaired in macrophages fromNOS2 knock-out mice (MacMicking et al 1997) Moreover animalstudies have demonstrated that in tumor vessels of melanomaxenografts macrophage-derived NO induced the expression of theadhesion molecule VCAM-1 which is important for T-cellextravasation Additionally only co-transfer of CD8+ T cells withwild-type macrophages but not withNos2minusminusmacrophages yieldedT-cell homing to the tumor and consequently led to tumor rejection(Sektioglu et al 2016)These studies collectively suggest that cellular levels of NO and
the associated cross-talk between cancer cells and theirenvironment are important for tumor initiation and progressionAs a consequence efforts have beenmade in recent years to improveNO detection and analysis (Box 2)
Metabolic regulation of NO synthesis in cancerCancer cells regulate the availability of intracellular arginine for NOsynthesis in multiple ways Cultured human colon cancer cells andmurine breast cancer cells stimulated by inflammatory mediatorssuch as LPS and IFN-γ increase the availability of arginine for NOproduction by enhancing transmembrane arginine import (Cendanet al 1996ab) Omental adipose stromal cells (O-ASCs) are
mesenchymal stem cells contained in the omentum tissue that areknown to promote endometrial and ovarian tumor proliferationInterestingly O-ASCs can support human endometrial or ovariancancer cells in co-culture by supplying them with arginine for NOproduction a finding that may reflect their role in tumor progression(Salimian Rizi et al 2015) Another way to upregulate argininelevels in tumor cells is to increase its endogenous synthesis Insamples from colon and breast cancer patients the overexpressionof ASL which encodes the enzyme that synthesizes arginineis associated with poor survival (Huang et al 2015 2017a)Additionally tumor samples from hepatocellular carcinoma patientsshow increased ASL expression and in human colon breast andhepatocellular carcinoma cells silencing of ASL expression byshort-hairpin RNA or the reduction of NO production by a NOSinhibitor inhibited cancer cellsrsquo proliferation and anchorage-independence (Box 1) (Huang et al 2013 2015 2017a) Thesestudies imply that ASL exerts its tumorigenic effects at least in partthrough NO but they do not rule out the possibility that increasedarginine levels might promote cancer by supporting the synthesis ofother arginine-derived molecules such as polyamines
Another urea cycle enzyme that functions in the arginine-citrulline cycle ASS1 is also overexpressed in various humancancers including in lung colon gastric and ovarian cancer(Delage et al 2010) The upregulated expression of ASS1 has beenfound to support the proliferation of human colon cancer cellsin vitro as well as the migration and metastatic potential of humangastric cell lines both in vitro and in mice with tumor xenografts(Bateman et al 2017 Shan et al 2015) Yet the cancer-promotingmechanisms fostered by ASS1 overexpression and their clinicalimplications remain unclear It is possible that high levels of ASS1support tumor proliferation and aggressiveness by increasing thesupply of arginine for NO production In support of this ASS1overexpression in rat vascular smooth muscle cells potentiated theLPS- and IFN-γ-stimulated production of NO (Xie and Gross1997) Moreover human breast cancer cells treated with the pro-inflammatory cytokine interleukin 17 (IL-17) increased theirproliferation and this response was reported to depend on the
Box 2 Detecting NO in cancerCellular concentrations of nitric oxide (NO) are hard to quantify due to itsvery short half-life In biological tissue samples NO levels can bemeasured indirectly by quantifying nitrite and nitrate using high-performance liquid chromatography (HPLC) (Jiang et al 2012) Thereagent 4-amino-5-methylamino-2prime7prime-difluorofluorescein (DAF-FM) isused for intracellular measurements because it becomes fluorescentwhen it reacts with NO and can be detected by any fluorescein-detectinginstrument (Namin et al 2013) Undoubtedly the ideal method formeasuring NO levels is via a flux analysis performed in vivo by usinglabeled arginine and then tracing the downstream products such ascitrulline and NO-derived urinary nitrate by using gas or liquidchromatography-mass spectrometry (Magneacute et al 2009)For a more global assessment of NO involvement in cancer the
expression of individual genes in the arginine-NO pathway can beassessed across different types of cancer in large datasets such as inThe Cancer Genome Atlas (TCGA) Expression levels can then becorrelated with patient survival and therapeutic response (Ekmekciogluet al 2016)Although these methods have all contributed to our understanding of
NO biology NO production and its downstream effects often involvecomplex signaling pathways and thus it remains challenging to dissectthe exact cellular contribution of NO to disease pathogenesis and toinvestigate its therapeutic relevance
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
decreasing the expression of the oncogene c-Myc in a cGMP-dependent manner (Ciani et al 2004) Moreover NO can inhibitthe proliferation of a human neuroblastoma cancer cell line in vitroby upregulating tumor suppressor pathways including the BRCA1Chk1p53 pathway leading to cell cycle arrest in response to DNAdamage via activation of cell cycle checkpoints (Van de Wouweret al 2012) Of note the dual roles of NO in cancer are also dose-dependent while exogenous NO stimulated cell proliferation inpheochromocytoma PC12 cells at low concentrations it inhibitedproliferation at higher concentrations (Bal-Price et al 2006)Likewise whereas low levels of NO can inhibit apoptosis andpromote cancer high levels of NO can contribute to cancer cellapoptosis (Villalobo 2006)NO has also been implicated in the epigenetic modification
of gene expression Several studies have described NO-drivenepigenetic modifications that control normal biologicaldevelopment and mediate tumorigenesis (Vasudevan et al 2016)In prostate carcinogenesis the silencing of glutathione transferaseP1-1 (GSTP1) is a common early event that is frequently caused bypromoter hypermethylation and correlates with decreased survivalIn human prostate cancer cells eNOS participates in GSTP1repression by being recruited to the gene promoter with aconsequential remodeling of the local chromatin (Re et al 2011)Importantly pharmacological inhibition of eNOS relieved therepression of GSTP1 and treatment with an NO donor silenced thisgene suggesting that eNOS regulates GSTP1 transcription throughNO production In oral squamous cell carcinoma patients it iscommon to find histone hyperacetylation that promotes tumorprogression Interestingly it has been found that NO mediateshistone hyperacetylation and that p300 histone acetylase activity isdependent on endogenously generated NO (Arif et al 2010)Moreover NO can affect histone PTMs at a global level astreatment with NO donors resulted in the differential expression of
over 6500 genes in breast cancer cells in which the pattern of PTMscorrelated with an oncogenic signature (Vasudevan et al 2015)Conversely NO has been found to inhibit lysine demethylase 3A(KDM3A) a histone demethylase that is known to positivelyregulate cancer cell invasion chemoresistance and metastasis inbreast and ovarian cancer cells (Hickok et al 2013)
The tumor microenvironment also influences the contribution ofNO to tumor fate Indeed cancer cells have developed diverse waysto intervene in the production andor metabolism of NO in tumorsand their surrounding tissue to gain an advantage As with itsdichotomous roles in cancer cell proliferation and apoptosis NOcan influence both cancer progression and its restriction NOS1upregulation was found to support the growth and activity ofcultured cancer-associated fibroblasts which are known to stimulatetumor progression (Augsten et al 2014) Additionally NO wasdemonstrated to directly promote cancer progression and invasionby influencing the stromal components of a tumor by inducingepithelial-to-mesenchymal transition (EMT Box 1) as well as byaffecting tumor vessel formation In human squamous cellcarcinoma and in lung cancer cells lines EMT and stem cellfeatures are reportedly activated by moderate levels of NO producedby NOS in response to growth factors and inflammatory mediators(Terzuoli et al 2017) On the other hand it seems that in the samecells higher than normal concentrations of NO inhibit EMT Inhuman metastatic prostate cancer cell lines treated with a highconcentration of the NO donor DETA-NONOate EMT and theinvasive phenotype of these cells is reversed via the inhibition of theEMT effector and transcription factor Snail (Baritaki et al 2010) Itwas demonstrated in this study that both Snail mRNA levels and itsDNA-binding capacity were inhibited by NO in a yet-undefinedmechanism Moreover NO was found to inhibit the mitochondrialfunction of Complex I and IV of the electron transport chain (Daiet al 2013) and to perturb the integrity of the mitochondrial
Growth factor induction
Oxidative DNA damage
Angiogenesis induction
Apoptosis inhibition
EMT induction
EMT inhibition
Apoptosis induction
Immune cell activation
Angiogenesis inhibition
Tumor suppressor pathway
activation
Cancer-promoting effects
Cancer-restrictingeffects
NO
Fig 2 Dichotomous roles for NO in cancer The dichotomousroles of NO in cancer are depicted in a Yin and Yang model toillustrate its cancer-promoting and cancer-restricting effectsNO can either inhibit or enhance cancer progression dependingon the biological context its concentration and on the duration ofNO production EMT epithelial-to-mesenchymal transitionNO nitric oxide
network by S-nitrosylation of dynamine-1-like protein (DRP-1)which is known to regulate mitochondrial fission (Cho et al 2009)It is thus tempting to speculate that the proliferation and EMTelicited by low doses of NO in cancer are associated with theinhibition of mitochondrial activity (Boland et al 2013)Cancer progression also depends on angiogenesis which is
needed to support the growing tumor with oxygen and nutrients andto remove waste products NO can promote angiogenesis bysupporting endothelial differentiation by inhibiting antiangiogenicfactors by dilating tumor blood vessels and by recruiting bone-marrow-derived and perivascular cells (Fukumura et al 2006) In amurine melanoma model lacking eNOS expression a perturbedrecruitment of mural cells to newly formed vessels and abnormalvessel branching and stabilization were demonstrated (Kashiwagiet al 2005) Moreover nNOS was required for the formationof abnormal tumor blood vessels in mice harboring humanglioma xenografts (Kashiwagi et al 2008) Conversely severalNO-related metabolites such as isosorbide mononitrate anddinitrate have been found to suppress VEGF protein levels incultured human colon cancer cells and to inhibit angiogenesisin vivo in xenografts of murine lung tumors (Pathi et al 2011Pipili-Synetos et al 1995)In recent years researchers have focused on the role of the
immune system in cancer development Here too NO produced byimmune cells has dual regulatory functions in tumor progressionNO and reactive nitrogen species that originate from immune cellssuch as macrophages and neutrophils can have pro-tumorigeniceffects on neighboring epithelial cells for example by inducingDNA damage that can initiate inflammation-associated neoplastictransformation (Wang et al 2017) NO produced by tumor-infiltrating myeloid cells was found to be important for activation ofadoptively transferred cytotoxic T cells (Marigo et al 2016) It wasalso shown that NO production by colon cells is required inpathogen-induced colon inflammation and immune cell infiltrationeventually leading to dysplasia and colon cancer development(Erdman et al 2009) In parallel NO can activate macrophages andcytotoxic T cells and augment the immune response against tumorcells (MacMicking et al 1997 Marigo et al 2016) Indeed thecytotoxic action of the cytokine interferon gamma (IFN-γ) or theactivity of LPS-activated primary mouse macrophages againstdifferent cancer cell lines were impaired in macrophages fromNOS2 knock-out mice (MacMicking et al 1997) Moreover animalstudies have demonstrated that in tumor vessels of melanomaxenografts macrophage-derived NO induced the expression of theadhesion molecule VCAM-1 which is important for T-cellextravasation Additionally only co-transfer of CD8+ T cells withwild-type macrophages but not withNos2minusminusmacrophages yieldedT-cell homing to the tumor and consequently led to tumor rejection(Sektioglu et al 2016)These studies collectively suggest that cellular levels of NO and
the associated cross-talk between cancer cells and theirenvironment are important for tumor initiation and progressionAs a consequence efforts have beenmade in recent years to improveNO detection and analysis (Box 2)
Metabolic regulation of NO synthesis in cancerCancer cells regulate the availability of intracellular arginine for NOsynthesis in multiple ways Cultured human colon cancer cells andmurine breast cancer cells stimulated by inflammatory mediatorssuch as LPS and IFN-γ increase the availability of arginine for NOproduction by enhancing transmembrane arginine import (Cendanet al 1996ab) Omental adipose stromal cells (O-ASCs) are
mesenchymal stem cells contained in the omentum tissue that areknown to promote endometrial and ovarian tumor proliferationInterestingly O-ASCs can support human endometrial or ovariancancer cells in co-culture by supplying them with arginine for NOproduction a finding that may reflect their role in tumor progression(Salimian Rizi et al 2015) Another way to upregulate argininelevels in tumor cells is to increase its endogenous synthesis Insamples from colon and breast cancer patients the overexpressionof ASL which encodes the enzyme that synthesizes arginineis associated with poor survival (Huang et al 2015 2017a)Additionally tumor samples from hepatocellular carcinoma patientsshow increased ASL expression and in human colon breast andhepatocellular carcinoma cells silencing of ASL expression byshort-hairpin RNA or the reduction of NO production by a NOSinhibitor inhibited cancer cellsrsquo proliferation and anchorage-independence (Box 1) (Huang et al 2013 2015 2017a) Thesestudies imply that ASL exerts its tumorigenic effects at least in partthrough NO but they do not rule out the possibility that increasedarginine levels might promote cancer by supporting the synthesis ofother arginine-derived molecules such as polyamines
Another urea cycle enzyme that functions in the arginine-citrulline cycle ASS1 is also overexpressed in various humancancers including in lung colon gastric and ovarian cancer(Delage et al 2010) The upregulated expression of ASS1 has beenfound to support the proliferation of human colon cancer cellsin vitro as well as the migration and metastatic potential of humangastric cell lines both in vitro and in mice with tumor xenografts(Bateman et al 2017 Shan et al 2015) Yet the cancer-promotingmechanisms fostered by ASS1 overexpression and their clinicalimplications remain unclear It is possible that high levels of ASS1support tumor proliferation and aggressiveness by increasing thesupply of arginine for NO production In support of this ASS1overexpression in rat vascular smooth muscle cells potentiated theLPS- and IFN-γ-stimulated production of NO (Xie and Gross1997) Moreover human breast cancer cells treated with the pro-inflammatory cytokine interleukin 17 (IL-17) increased theirproliferation and this response was reported to depend on the
Box 2 Detecting NO in cancerCellular concentrations of nitric oxide (NO) are hard to quantify due to itsvery short half-life In biological tissue samples NO levels can bemeasured indirectly by quantifying nitrite and nitrate using high-performance liquid chromatography (HPLC) (Jiang et al 2012) Thereagent 4-amino-5-methylamino-2prime7prime-difluorofluorescein (DAF-FM) isused for intracellular measurements because it becomes fluorescentwhen it reacts with NO and can be detected by any fluorescein-detectinginstrument (Namin et al 2013) Undoubtedly the ideal method formeasuring NO levels is via a flux analysis performed in vivo by usinglabeled arginine and then tracing the downstream products such ascitrulline and NO-derived urinary nitrate by using gas or liquidchromatography-mass spectrometry (Magneacute et al 2009)For a more global assessment of NO involvement in cancer the
expression of individual genes in the arginine-NO pathway can beassessed across different types of cancer in large datasets such as inThe Cancer Genome Atlas (TCGA) Expression levels can then becorrelated with patient survival and therapeutic response (Ekmekciogluet al 2016)Although these methods have all contributed to our understanding of
NO biology NO production and its downstream effects often involvecomplex signaling pathways and thus it remains challenging to dissectthe exact cellular contribution of NO to disease pathogenesis and toinvestigate its therapeutic relevance
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
network by S-nitrosylation of dynamine-1-like protein (DRP-1)which is known to regulate mitochondrial fission (Cho et al 2009)It is thus tempting to speculate that the proliferation and EMTelicited by low doses of NO in cancer are associated with theinhibition of mitochondrial activity (Boland et al 2013)Cancer progression also depends on angiogenesis which is
needed to support the growing tumor with oxygen and nutrients andto remove waste products NO can promote angiogenesis bysupporting endothelial differentiation by inhibiting antiangiogenicfactors by dilating tumor blood vessels and by recruiting bone-marrow-derived and perivascular cells (Fukumura et al 2006) In amurine melanoma model lacking eNOS expression a perturbedrecruitment of mural cells to newly formed vessels and abnormalvessel branching and stabilization were demonstrated (Kashiwagiet al 2005) Moreover nNOS was required for the formationof abnormal tumor blood vessels in mice harboring humanglioma xenografts (Kashiwagi et al 2008) Conversely severalNO-related metabolites such as isosorbide mononitrate anddinitrate have been found to suppress VEGF protein levels incultured human colon cancer cells and to inhibit angiogenesisin vivo in xenografts of murine lung tumors (Pathi et al 2011Pipili-Synetos et al 1995)In recent years researchers have focused on the role of the
immune system in cancer development Here too NO produced byimmune cells has dual regulatory functions in tumor progressionNO and reactive nitrogen species that originate from immune cellssuch as macrophages and neutrophils can have pro-tumorigeniceffects on neighboring epithelial cells for example by inducingDNA damage that can initiate inflammation-associated neoplastictransformation (Wang et al 2017) NO produced by tumor-infiltrating myeloid cells was found to be important for activation ofadoptively transferred cytotoxic T cells (Marigo et al 2016) It wasalso shown that NO production by colon cells is required inpathogen-induced colon inflammation and immune cell infiltrationeventually leading to dysplasia and colon cancer development(Erdman et al 2009) In parallel NO can activate macrophages andcytotoxic T cells and augment the immune response against tumorcells (MacMicking et al 1997 Marigo et al 2016) Indeed thecytotoxic action of the cytokine interferon gamma (IFN-γ) or theactivity of LPS-activated primary mouse macrophages againstdifferent cancer cell lines were impaired in macrophages fromNOS2 knock-out mice (MacMicking et al 1997) Moreover animalstudies have demonstrated that in tumor vessels of melanomaxenografts macrophage-derived NO induced the expression of theadhesion molecule VCAM-1 which is important for T-cellextravasation Additionally only co-transfer of CD8+ T cells withwild-type macrophages but not withNos2minusminusmacrophages yieldedT-cell homing to the tumor and consequently led to tumor rejection(Sektioglu et al 2016)These studies collectively suggest that cellular levels of NO and
the associated cross-talk between cancer cells and theirenvironment are important for tumor initiation and progressionAs a consequence efforts have beenmade in recent years to improveNO detection and analysis (Box 2)
Metabolic regulation of NO synthesis in cancerCancer cells regulate the availability of intracellular arginine for NOsynthesis in multiple ways Cultured human colon cancer cells andmurine breast cancer cells stimulated by inflammatory mediatorssuch as LPS and IFN-γ increase the availability of arginine for NOproduction by enhancing transmembrane arginine import (Cendanet al 1996ab) Omental adipose stromal cells (O-ASCs) are
mesenchymal stem cells contained in the omentum tissue that areknown to promote endometrial and ovarian tumor proliferationInterestingly O-ASCs can support human endometrial or ovariancancer cells in co-culture by supplying them with arginine for NOproduction a finding that may reflect their role in tumor progression(Salimian Rizi et al 2015) Another way to upregulate argininelevels in tumor cells is to increase its endogenous synthesis Insamples from colon and breast cancer patients the overexpressionof ASL which encodes the enzyme that synthesizes arginineis associated with poor survival (Huang et al 2015 2017a)Additionally tumor samples from hepatocellular carcinoma patientsshow increased ASL expression and in human colon breast andhepatocellular carcinoma cells silencing of ASL expression byshort-hairpin RNA or the reduction of NO production by a NOSinhibitor inhibited cancer cellsrsquo proliferation and anchorage-independence (Box 1) (Huang et al 2013 2015 2017a) Thesestudies imply that ASL exerts its tumorigenic effects at least in partthrough NO but they do not rule out the possibility that increasedarginine levels might promote cancer by supporting the synthesis ofother arginine-derived molecules such as polyamines
Another urea cycle enzyme that functions in the arginine-citrulline cycle ASS1 is also overexpressed in various humancancers including in lung colon gastric and ovarian cancer(Delage et al 2010) The upregulated expression of ASS1 has beenfound to support the proliferation of human colon cancer cellsin vitro as well as the migration and metastatic potential of humangastric cell lines both in vitro and in mice with tumor xenografts(Bateman et al 2017 Shan et al 2015) Yet the cancer-promotingmechanisms fostered by ASS1 overexpression and their clinicalimplications remain unclear It is possible that high levels of ASS1support tumor proliferation and aggressiveness by increasing thesupply of arginine for NO production In support of this ASS1overexpression in rat vascular smooth muscle cells potentiated theLPS- and IFN-γ-stimulated production of NO (Xie and Gross1997) Moreover human breast cancer cells treated with the pro-inflammatory cytokine interleukin 17 (IL-17) increased theirproliferation and this response was reported to depend on the
Box 2 Detecting NO in cancerCellular concentrations of nitric oxide (NO) are hard to quantify due to itsvery short half-life In biological tissue samples NO levels can bemeasured indirectly by quantifying nitrite and nitrate using high-performance liquid chromatography (HPLC) (Jiang et al 2012) Thereagent 4-amino-5-methylamino-2prime7prime-difluorofluorescein (DAF-FM) isused for intracellular measurements because it becomes fluorescentwhen it reacts with NO and can be detected by any fluorescein-detectinginstrument (Namin et al 2013) Undoubtedly the ideal method formeasuring NO levels is via a flux analysis performed in vivo by usinglabeled arginine and then tracing the downstream products such ascitrulline and NO-derived urinary nitrate by using gas or liquidchromatography-mass spectrometry (Magneacute et al 2009)For a more global assessment of NO involvement in cancer the
expression of individual genes in the arginine-NO pathway can beassessed across different types of cancer in large datasets such as inThe Cancer Genome Atlas (TCGA) Expression levels can then becorrelated with patient survival and therapeutic response (Ekmekciogluet al 2016)Although these methods have all contributed to our understanding of
NO biology NO production and its downstream effects often involvecomplex signaling pathways and thus it remains challenging to dissectthe exact cellular contribution of NO to disease pathogenesis and toinvestigate its therapeutic relevance
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
enhanced availability of arginine for NO production This increasedarginine flux was associated with the upregulated expression of bothASS1 andNOS3 and with the downregulated expression of arginase(Amara et al 2017)ASS1 expression can also be downregulated in some cancers in
association with the methylation of its promoter (Wu et al 2013)Reduced ASS1 expression has been associated with higherrecurrence shorter disease-free survival and with shorter overallsurvival in patients with pancreatic cancer (Liu et al 2017) Inosteosarcoma patients lower ASS1 expression levels in tumorsamples were associated with resistance to doxorubicin treatment(Kim et al 2016) and with the development of pulmonarymetastases (Kobayashi et al 2010) Interestingly ASS1 and ASLexpression was silenced by gene promoter methylation in primarycultures of human glioblastoma multiforme cells suggesting thatthese genes are not mutually exclusive and hence silencing of eachone of these genes may modulate a separate metabolic pathway(Syed et al 2013) Indeed independent of arginine importancecancer cells become more proliferative when ASS1 is silencedbecause of the increased cytosolic availability of its substrateaspartate for pyrimidine synthesis by CAD (carbamoyl-phosphatesynthase 2 aspartate transcarbamylase and dihydroorotase)(Rabinovich et al 2015 Moreno-Morcillo et al 2017)Nevertheless it is also possible that ASS1 downregulationpromotes cancer by decreasing the availability of arginine for NOsynthesis Importantly the silencing of ASS1 or ASL in tumorsresults in arginine auxotrophy ndash an intrinsic dependence of the cellson exogenous arginine due to their inability to synthesize it In thesecircumstances arginine becomes an essential amino acidgenerating a vulnerability that can be used to treat cancer usingarginine-depriving agents (Syed et al 2013)Cancer cells can also increase NO production via the
upregulation of NOS Indeed it has been demonstrated thatfollowing the exposure of a human osteosarcoma cell line to LPS- orIFN-γ-induced iNOS NO levels subsequently increased(Tachibana et al 2000) Moreover hypoxia and inflammatorycytokines can induce iNOS expression in cultured human breastcancer cells with a subsequent elevation in poor-survivalbiomarkers such as S100 calcium-binding protein A8 IL-6 IL-8and tissue inhibitor of matrix metalloproteinase-1 (Heinecke et al2014) In addition iNOSmRNA and protein levels were found to beelevated in tumor samples from patients with nasopharyngealcarcinoma (Segawa et al 2008) Another interesting cancer-associated mechanism involving NOS is the reduction in theavailability of BH4 which has been reported in human breastcolorectal epidermoid and head and neck tumors as compared tonormal human tissues (Rabender et al 2015) As a consequencerather than NO NOS activity generates more peroxynitrite whichhas anti-apoptotic signaling properties (Delgado-Esteban et al2007) Accordingly treating human breast cancer cells with a BH4precursor inhibited their growth both in culture and in tumorxenografts in vivo (Rabender et al 2015) In addition to NOproduced by cytosolic NOS NO produced by the mitochondrialNOS has also been found to be relevant to cancer MitochondrialNO synthesis requires at least in part the transport of arginine to themitochondria through the solute carrier family 25 member 29(SLC25A29) transporter (Porcelli et al 2014) The relevance ofthis diversion of arginine to mitochondrial NO synthesis isillustrated by the deleterious consequences of SLC25A29deletion in cancer cells which was found to impair NOproduction and to reduce tumor growth (Zhang et al 2018)However the regulation and molecular consequences of the
cytoplasmic versus mitochondrial production of NO requirefurther investigation
The enzyme ARG2 competes with NOS for arginine as asubstrate accordingly as mentioned above the downregulation ofARG2 might increase the availability of arginine for NO production(Amara et al 2017) In contrast human breast tumor tissues expresshigh levels of the ARG2 gene and its inhibition in breast cancer cellxenografts by the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMGCoA) reductase inhibitor rosuvastatin led to the inhibition of tumorproliferation (Erbas et al 2015 Singh et al 2013) This therapeuticeffect was suggested to be due to a reduction in the use of arginine asa precursor for polyamines with a concomitant increase in its usageas a precursor for NO (Cervelli et al 2014)
NO-related drugs for cancer therapyBecause high NO levels have been shown to play a tumorigenic rolein various types of cancer one rational approach to treating suchcancers is to develop drugs that decrease NO levels (Fig 3) Severaldrugs that inhibit NOS enzymatic activity exist (Paige and Jaffrey2007) However clinical trial results were complex For exampleNOS inhibition using drugs such as Sanggenon C or L-NG-nitroarginine methyl ester (L-NAME) restricted tumor growth inmouse xenograft cancer models (Chen et al 2017 Pershing et al2016 Ridnour et al 2015) Yet a Phase 1 clinical trial of the iNOSinhibitor ASP9853 used in combination with the chemotherapeuticdrug docetaxel to treat patients with resistant solid tumors wasprematurely terminated because of neutropenia-associated toxicities(Luke et al 2016) Interestingly the endogenous compoundasymmetric dimethylarginine (ADMA) which inhibits NOS bycompeting with arginine has recently been shown to be degradedby the enzyme dimethylarginine dimethylaminohydrolase-1(DDAH1) DDAH1 is frequently upregulated in prostate cancerwhere it promotes tumor growth and angiogenesis suggesting thatanti-cancer drugs that induce ADMA or that inhibit DDAH1 couldpotentially be useful in treating tumors that are influenced by thepro-tumorigenic properties of NO (Reddy et al 2018)
As discussed above arginine-depleting agents are being tested astreatment for tumors that are auxotrophic for arginine The enzymearginine deiminase (ADI) which allows many microorganisms toutilize arginine as a major energy source was recently included inclinical trials as an anti-cancer drug to treat arginine-auxotrophictumors with positive effects reported on reducing diseaseprogression in hepatocellular carcinoma advanced pancreaticadenocarcinoma and acute myeloid leukemia patients (Izzo et al2004 Lowery et al 2017 Tsai et al 2017) Since NO is animportant product of arginine in cancer cells it is plausible thatarginine depletion might contribute to tumor inhibition by reducingthe cellular levels of NO In Phase 1 and 2 clinical trials metastaticmelanoma patients responded to ADI treatment and showed reducedplasma NO levels however no causative effect was proven betweenplasma NO levels and the clinical response to treatment (Asciertoet al 2005) Another arginine-depleting agent a PEGylatedderivative of recombinant human ARG1 was found to inhibitcancer progression when tested in a Phase 1 clinical trial for thetreatment of hepatocellular carcinoma patients (Yau et al 2013)
In addition to decreasing NO as a form of anti-cancer therapyelevating NO to cytotoxic levels using NO donors has also beentried therapeutically NO donors can potentially exert their anti-tumor activity by acting directly to reduce cancer progression butalso indirectly by increasing tumor blood flow to enhance thedelivery of cytotoxic therapy to tumor tissue (Ning et al 2014)Notably the use of NO donors as a form of anti-cancer therapy
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
has been challenging due to the short half-life of NO and theneed to specifically target the right cells with the right doseThus various NO donors are being tested in clinical trials as anti-cancer therapeutics in combination with either chemotherapyradiotherapy or immunotherapy to overcome NO-related treatmentdifficulties as well as tumor cell resistance to conventionaltreatments (Huang et al 2017b) Encouragingly NO donorsinhibit cultured human ovarian cancer cell survival and anti-apoptotic pathways such as the NF-κB signaling cascade (knownphysiologically to be negatively regulated by S-nitrosylation) andsensitize drug-resistant tumor cells to apoptosis by bothchemotherapy and immunotherapy (Bonavida et al 2008 Garbanand Bonavida 2001 Marshall and Stamler 2001) The NO donorglyceryl tri-nitrate has also been found to inhibit tumor progressionin a Phase 2 study of prostate cancer patients following primarytreatment failure (Siemens et al 2009) Glyceryl tri-nitrate is also apromising chemo-sensitizing agent for advanced non-small-celllung cancer (Dingemans et al 2015) and for advanced rectalcancer when used in combination with chemotherapy andradiotherapy (Illum et al 2015) In another Phase 2 clinical trialpretreatment of refractory lung cancer patients with the NO
donor 1-bromoacetyl-33-dinitroazetidine (RRx-001) sensitized thepatients to the chemotherapeutic drug carboplatin (Carter et al2016) Combining NO donors with other agents or anti-cancerdrugs is another strategy that has proved to be effectiveagainst various cancer cell lines in preclinical models (Huang et al2017b) NO coupled to a non-steroidal anti-inflammatory molecule(NO-NSAID) induced apoptosis and modulated Wnt and NF-κBsignaling in human colon cancer cells in vitro (Rigas and Kashfi2004) as well as in vivo in a breast cancer mouse model (Nathet al 2015)
The mobilization of the immune system to treat cancer has taken acentral stage in cancer therapy in recent years A recent studydemonstrated that hypoxia-induced expression of the immuneinhibitory molecule programmed cell death ligand-1 (PD-L1) inmurine melanoma cancer cells increased their resistance to lysis byin-vivo-generated cytotoxic T lymphocytes (CTLs) in a hypoxia-inducible factor-1α (HIF-1α)-dependent manner (Barsoum et al2014) Notably as NO signaling activation was previously shown toprevent hypoxia-induced accumulation of HIF-1α (Barsoum et al2011) treatment with the NO donor glyceryl tri-nitrate preventedthe hypoxia-induced expression of PDL1 in murine melanoma
Arginine
Ornithine
Citrulline
ASA
Fumarate
ine
Aspartate
NOS
ASS1
NO
y+ transport sy
stem
Cytosol
Arginine-citrulline
cycle
Arginine
ASL
Arginase 2
Pyrimidines
Polyamines
Elevation of NO levels byNO donors
NOSinhibitors
PEGylated ADIPEGylated
arginase
Fig 3 NO-metabolism-related anti-cancer strategies A schematic illustration of NO metabolic pathways in a cancer cell Red arrows denote the cancer-related up- or down-regulation of proteins that are involved in NOmetabolism leading to a net increase in NO production Tumor cells can enhance NO productionby upregulating NOS levels increasing arginine transport increasing the levels of ASS1 and ASL to enhance arginine availability for NO synthesis or bydecreasing arginine metabolism by inhibiting arginase Purple arrows denote the cancer-related up- or down-regulation of proteins involved in NO metabolismleading to a net decrease in NO production In addition to restricting NO levels ASS1 inhibition and ARG2 upregulationmight alsometabolically support cancer byincreasing the production of pyrimidines and polyamines respectively (dashed blue arrows) Anti-cancer NO-related strategies that increase NO levels aredenoted in a red box and those that downregulate NO levels are depicted in blue boxes NO-related anti-cancer strategies include increasing levels of NOwith NOdonors or decreasing NO levels via NOS inhibition or with PEGylated arginine-degrading enzymes ADI arginine deiminase ASA argininosuccinic acid ASLargininosuccinate lyase ASS1 argininosuccinate synthase 1 NO nitric oxide NOS nitric oxide synthase
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
cancer cells and diminished the cellsrsquo resistance to CTL-mediatedlysis indicating the potential use of NO donors asimmunotherapeutic drugs against hypoxic tumor cells Howevertreating hypoxic tumors with NO may be challenging as NO wasalso demonstrated to enhance HIF-1α levels and activity in culturedhuman cancer cells (Berchner-Pfannschmidt et al 2007 Kimuraet al 2000 Thomas et al 2004)
ConclusionsSince its discovery more than 200 years ago numerous studies haveidentified NO as an important cellular signaling molecule involvedin many physiological and pathological processes Not surprisinglyNO is also emerging as a central player in cancer due to itscontribution to tumor initiation and progression However thepleiotropic nature of its physiological roles its complicated spatialtemporal and dosage regulation at multiple levels together with itsshort half-life make NO challenging to target therapeuticallyIndeed a cell-specific approach is required to induce or inhibit NOsynthesis for cancer therapy Encouragingly NO can also beregulated metabolically through the availability of arginine makingarginine a potential therapeutic target Indeed the activity of the keyenzymes involved in NO production namely ASS1 ASL arginaseand NOS are frequently altered in various types of cancersenabling us to identify vulnerabilities in NO-related pathways and todesign novel anti-cancer drugs that target these enzymes Recentlystudies showed that the metabolic supplementation of citrullinewhich drives NO synthesis (Kim et al 2015) together with fisetinwhich upregulates ASL and ASS1 levels is a promising approachto overcoming NO-related tissue and dosage obstacles and torestrict the development of inflammation-associated colon cancer(Stettner et al 2018) In this approach the body diverts themetabolic supplements to enable NO synthesis at the right placeand at the required dosage Undoubtedly further advances in ourunderstanding of the pathology of different cancers and in thetechniques for detecting NO will strengthen our understandingof the ways in which arginine-NO metabolism contributes tocancer and will aid in the development of related anti-cancertherapeutic approaches
This article is part of a special subject collection lsquoCancer Metabolism modelsmechanisms and targetsrsquo which was launched in a dedicated issue guest edited byAlmut Schulze and Mariia Yuneva See related articles in this collection at httpdmm biologistsorgcollectioncancermetabolism
Competing interestsThe authors declare no competing or financial interests
FundingAE is the incumbent of the Leah Omenn Career Development Chair and issupported by research grants from the European Research Council Program(CIG618113 ERC614204) the Israel Science Foundation (134313 195213) and aMinerva Foundation grant award (711730) AE received additional support from theAdelis Foundation the Henry S and Anne S Reich Research Fund the Dukler Fundfor Cancer Research the Paul Sparr Foundation the Saul and Theresa EsmanFoundation from Joseph Piko Baruch and from the estate of Fannie Sherr RK issupported by the Rising Tide Foundation (RTF) fellowship
ReferencesAdams L Franco M C and Estevez A G (2015) Reactive nitrogen species incellular signaling Exp Biol Med 240 711-717
Amara S Majors C Roy B Hill S Rose K L Myles E L and Tiriveedhi V(2017) Critical role of SIK3 in mediating high salt and IL-17 synergy leading tobreast cancer cell proliferation PLoS ONE 12 e0180097
Arancibia-Garavilla Y Toledo F Casanello P and Sobrevia L (2003) Nitricoxide synthesis requires activity of the cationic and neutral amino acid transportsystem y+L in human umbilical vein endothelium Exp Physiol 88 699-710
Arif M Vedamurthy B M Choudhari R Ostwal Y B Mantelingu KKodaganur G S and Kundu T K (2010) Nitric oxide-mediated histonehyperacetylation in oral cancer target for a water-soluble HAT inhibitor CTK7AChem Biol 17 903-913
Ascierto P A Scala S Castello G Daponte A Simeone E Ottaiano ABeneduce G De Rosa V Izzo F Melucci M T et al (2005) Pegylatedarginine deiminase treatment of patients with metastatic melanoma results fromphase I and II studies J Clin Oncol 23 7660-7668
Augsten M Sjoberg E Frings O Vorrink S U Frijhoff J Olsson E BorgA and Ostman A (2014) Cancer-associated fibroblasts expressing CXCL14rely upon NOS1-derived nitric oxide signaling for their tumor-supportingproperties Cancer Res 74 2999-3010
Bal-Price A Gartlon J and Brown G C (2006) Nitric oxide stimulates PC12cell proliferation via cGMP and inhibits at higher concentrations mainly via energydepletion Nitric Oxide 14 238-246
Baritaki S Huerta-Yepez S Sahakyan A Karagiannides I Bakirtzi KJazirehi A and Bonavida B (2010) Mechanisms of nitric oxide-mediatedinhibition of EMT in cancer inhibition of the metastasis-inducer Snail andinduction of the metastasis-suppressor RKIP Cell Cycle 9 4931-4940
Barsoum I B Hamilton T K Li X Cotechini T Miles E A Siemens D Rand Graham C H (2011) Hypoxia induces escape from innate immunity incancer cells via increased expression of ADAM10 role of nitric oxideCancer Res71 7433-7441
Barsoum I B Smallwood C A Siemens D R and Graham C H (2014) Amechanism of hypoxia-mediated escape from adaptive immunity in cancer cellsCancer Res 74 665-674
Bateman L A Ku W-M Heslin M J Contreras C M Skibola C F andNomura D K (2017) Argininosuccinate synthase 1 is a metabolic regulator ofcolorectal cancer pathogenicity ACS Chem Biol 12 905-911
Berchner-Pfannschmidt U Yamac H Trinidad B and Fandrey J (2007)Nitric oxide modulates oxygen sensing by hypoxia-inducible factor 1-dependentinduction of prolyl hydroxylase 2 J Biol Chem 282 1788-1796
Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity anupdate Trends Immunol 36 161-178
Boland M L Chourasia A H and Macleod K F (2013) MitochondrialDysfunction in Cancer Front Oncol 3 292
Bonavida B Baritaki S Huerta-Yepez S Vega M I Chatterjee D andYeung K (2008) Novel therapeutic applications of nitric oxide donors in cancerroles in chemo- and immunosensitization to apoptosis and inhibition ofmetastases Nitric Oxide 19 152-157
Bredt D S (1999) Endogenous nitric oxide synthesis biological functions andpathophysiology Free Radic Res 31 577-596
Burke A J Sullivan F J Giles F J and Glynn S A (2013) The yin and yangof nitric oxide in cancer progression Carcinogenesis 34 503-512
Caldwell R B Toque H A Narayanan S P and Caldwell R W (2015)Arginase an old enzyme with new tricks Trends Pharmacol Sci 36 395-405
Carter C A Oronsky B Caroen S Scicinski J Cabrales P Degesys Aand Brzezniak C (2016) Partial response to carboplatin in an RRx-001pretreated patient with EGFR-inhibitor-resistance and T790M-negative NSCLCRespir Med Case Rep 18 62-65
Cendan J C Souba W W Copeland E M III and Lind D S (1996a)Increased L-arginine transport in a nitric oxide-producing metastatic colon cancercell line Ann Surg Oncol 3 501-508
Cendan J C Topping D L Pruitt J Snowdy S Copeland E M III andLind D S (1996b) Inflammatory mediators stimulate arginine transport andarginine-derived nitric oxide production in a murine breast cancer cell line J SurgRes 60 284-288
Cervelli M Pietropaoli S Signore F Amendola R and Mariottini P (2014)Polyamines metabolism and breast cancer state of the art and perspectivesBreast Cancer Res Treat 148 233-248
Chen L-D Liu Z-H Zhang L-F Yao J-N and Wang C-F (2017)Sanggenon C induces apoptosis of colon cancer cells via inhibition of NOproduction iNOS expression and ROS activation of the mitochondrial pathwayOncol Rep 38 2123-2131
Cho D-H Nakamura T Fang J Cieplak P Godzik A Gu Z and LiptonS A (2009) S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrialfission and neuronal injury Science 324 102-105
Chuaiphichai S Crabtree M J McNeill E Hale A B Trelfa L ChannonK M and Douglas G (2017) A key role for tetrahydrobiopterin-dependentendothelial NOS regulation in resistance arteries studies in endothelial celltetrahydrobiopterin-deficient mice Br J Pharmacol 174 657-671
Ciani E Severi S Contestabile A Bartesaghi R and Contestabile A(2004) Nitric oxide negatively regulates proliferation and promotes neuronaldifferentiation through N-Myc downregulation J Cell Sci 117 4727-4737
Connelly L Jacobs A T Palacios-Callender M Moncada S and HobbsA J (2003) Macrophage endothelial nitric-oxide synthase autoregulates cellularactivation and pro-inflammatory protein expression J Biol Chem 27826480-26487
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
Dai Z Wu Z Yang Y Wang J Satterfield M C Meininger C J BazerF W and Wu G (2013) Nitric oxide and energy metabolism in mammalsBiofactors 39 383-391
Delage B Fennell D A Nicholson L McNeish I Lemoine N R Crook Tand Szlosarek P W (2010) Arginine deprivation and argininosuccinatesynthetase expression in the treatment of cancer Int J Cancer 126 2762-2772
Delgado-Esteban M Martin-Zanca D Andres-Martin L Almeida A andBolanos J P (2007) Inhibition of PTEN by peroxynitrite activates thephosphoinositide-3-kinaseAkt neuroprotective signaling pathwayJ Neurochem 102 194-205
Dingemans A-M C Groen H J M Herder G J M Stigt J A Smit E FBahce I Burgers J A van den Borne B E E M Biesma B Vincent Aet al (2015) A randomized phase II study comparing paclitaxel-carboplatin-bevacizumab with or without nitroglycerin patches in patients with stage IVnonsquamous nonsmall-cell lung cancer NVALT12 (NCT01171170)daggerAnnOncol 26 2286-2293
Dioguardi F S (2011) To give or not to give Lessons from the arginine paradoxJ Nutrigenet Nutrigenomics 4 90-98
Doulias P-T Tenopoulou M Greene J L Raju K and Ischiropoulos H(2013) Nitric oxide regulates mitochondrial fatty acid metabolism throughreversible protein S-nitrosylation Sci Signal 6 rs1
Du Q Zhang X Liu Q Zhang X Bartels C E and Geller D A (2013) Nitricoxide production upregulates Wntbeta-catenin signaling by inhibiting Dickkopf-1Cancer Res 73 6526-6537
Ekmekcioglu S Davies M A Tanese K Roszik J Shin-Sim M BassettR L Jr Milton D R Woodman S E Prieto V G Gershenwald J E et al(2016) Inflammatory marker testing identifies CD74 expression in melanomatumor cells and its expression associates with favorable survival for stage IIImelanoma Clin Cancer Res 22 3016-3024
Erbas H Bal O and Cakir E (2015) Effect of rosuvastatin on arginase enzymeactivity and polyamine production in experimental breast cancer Balkan Med J32 89-95
Erdman S E Rao V P Poutahidis T Rogers A B Taylor C L JacksonE A Ge Z Lee C W Schauer D B Wogan G N et al (2009) Nitric oxideand TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacterhepaticus-infected Rag2-deficient mice Proc Natl Acad Sci USA 1061027-1032
Fraisl P (2013) Crosstalk between oxygen- and nitric oxide-dependent signalingpathways in angiogenesis Exp Cell Res 319 1331-1339
Fukumura D Kashiwagi S and Jain R K (2006) The role of nitric oxide intumour progression Nat Rev Cancer 6 521-534
Garban H J and Bonavida B (2001) Nitric oxide disrupts H2O2-dependentactivation of nuclear factor kappa B Role in sensitization of human tumor cells totumor necrosis factor-alpha -induced cytotoxicity J Biol Chem 276 8918-8923
Garrido P Shalaby A Walsh E M Keane N Webber M Keane M MSullivan F J Kerin M J Callagy G Ryan A E et al (2017) Impact ofinducible nitric oxide synthase (iNOS) expression on triple negative breast canceroutcome and activation of EGFR and ERK signaling pathways Oncotarget 880568-80588
Ghafourifar P and Richter C (1997) Nitric oxide synthase activity inmitochondria FEBS Lett 418 291-296
Gould N Doulias P-T Tenopoulou M Raju K and Ischiropoulos H (2013)Regulation of protein function and signaling by reversible cysteine S-nitrosylationJ Biol Chem 288 26473-26479
Heinecke J L Ridnour L A Cheng R Y S Switzer C H Lizardo M MKhanna C Glynn S A Hussain S P Young H A Ambs S et al (2014)Tumormicroenvironment-based feed-forward regulation of NOS2 in breast cancerprogression Proc Natl Acad Sci USA 111 6323-6328
Hess D T Matsumoto A Kim S-O Marshall H E and Stamler J S (2005)Protein S-nitrosylation purview and parameters Nat Rev Mol Cell Biol 6150-166
Hickok J R Vasudevan D Antholine W E and Thomas D D (2013) Nitricoxide modifies global histone methylation by inhibiting Jumonji C domain-containing demethylases J Biol Chem 288 16004-16015
Huang H-L Hsu H-P Shieh S-C Chang Y-S Chen W-C Cho C-YTeng C-F Su I-J Hung W-C and Lai M-D (2013) Attenuation ofargininosuccinate lyase inhibits cancer growth via cyclin A2 and nitric oxide MolCancer Ther 12 2505-2516
Huang H L ChenW C Hsu H P Cho C Y Hung Y HWang C Y and LaiM D (2015) Argininosuccinate lyase is a potential therapeutic target in breastcancer Oncol Rep 34 3131-3139
Huang H-L Chen W-C Hsu H-P Cho C-Y Hung Y-H Wang C-Y andLai M-D (2017a) Silencing of argininosuccinate lyase inhibits colorectal cancerformation Oncol Rep 37 163-170
Huang Z Fu J and Zhang Y (2017b) Nitric oxide donor-based cancer therapyadvances and prospects J Med Chem 60 7617-7635
Husson A Brasse-Lagnel C Fairand A Renouf S and Lavoinne A (2003)Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycleEur J Biochem 270 1887-1899
Illum H Wang D H Dowell J E Hittson W J Torrisi J R Meyer J andHuerta S (2015) Phase I dose escalation trial of nitroglycerin in addition to 5-fluorouracil and radiation therapy for neoadjuvant treatment of operable rectalcancer Surgery 158 460-465
Izzo F Marra P Beneduce G Castello G Vallone P De Rosa VCremona F Ensor C M Holtsberg F W Bomalaski J S et al (2004)Pegylated arginine deiminase treatment of patients with unresectablehepatocellular carcinoma results from phase III studies J Clin Oncol 221815-1822
Jiang H Parthasarathy D Torregrossa A C Mian A and Bryan N S(2012) Analytical techniques for assaying nitric oxide bioactivity J Vis Exp 18e3722
Kashiwagi S Izumi Y Gohongi T Demou Z N Xu L Huang P L BuerkD G Munn L L Jain R K and Fukumura D (2005) NOmediates mural cellrecruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels J Clin Invest 115 1816-1827
Kashiwagi S Tsukada K Xu L Miyazaki J Kozin S V Tyrrell J ASessaW C Gerweck L E Jain R K and Fukumura D (2008) Perivascularnitric oxide gradients normalize tumor vasculature Nat Med 14 255-257
Kim I-Y Schutzler S E Schrader A Spencer H J Azhar G DeutzN E P andWolfe R R (2015) Acute ingestion of citrulline stimulates nitric oxidesynthesis but does not increase blood flow in healthy young and older adults withheart failure Am J Physiol Endocrinol Metab 309 E915-E924
Kim Y Kobayashi E Kubota D Suehara Y Mukaihara K Akaike K ItoA Kaneko K Chuman H Kawai A et al (2016) Reduced argininosuccinatesynthetase expression in refractory sarcomas impacts on therapeutic potentialand drug resistance Oncotarget 7 70832-70844
Kimura H Weisz A Kurashima Y Hashimoto K Ogura T DrsquoAcquisto FAddeo R Makuuchi M and Esumi H (2000) Hypoxia response element ofthe human vascular endothelial growth factor gene mediates transcriptionalregulation by nitric oxide control of hypoxia-inducible factor-1 activity by nitricoxide Blood 95 189-197
Kobayashi E Masuda M Nakayama R Ichikawa H Satow R ShitashigeM Honda K Yamaguchi U Shoji A Tochigi N et al (2010) Reducedargininosuccinate synthetase is a predictive biomarker for the development ofpulmonary metastasis in patients with osteosarcoma Mol Cancer Ther 9535-544
Kong Y Grimaldi M Curtin E Dougherty M Kaufman C White R MZon L I and Liao E C (2014) Neural crest development and craniofacialmorphogenesis is coordinated by nitric oxide and histone acetylation Chem Biol21 488-501
Liang X Zhang L Natarajan S K and Becker D F (2013) Prolinemechanisms of stress survival Antioxid Redox Signal 19 998-1011
Lind M Hayes A Caprnda M Petrovic D Rodrigo L Kruzliak P and ZulliA (2017) Inducible nitric oxide synthase good or bad Biomed Pharmacother93 370-375
Liu Q Stewart J Wang H Rashid A Zhao J Katz M H Lee J EFleming J B Maitra A Wolff R A et al (2017) Reduced expression ofargininosuccinate synthetase 1 has a negative prognostic impact in patients withpancreatic ductal adenocarcinoma PLoS ONE 12 e0171985
Lopez-Rivera E Jayaraman P Parikh F Davies M A Ekmekcioglu SIzadmehr S Milton D R Chipuk J E Grimm E A Estrada Y et al(2014) Inducible nitric oxide synthase drives mTOR pathway activation andproliferation of humanmelanoma by reversible nitrosylation of TSC2Cancer Res74 1067-1078
Lowery M A Yu K H Kelsen D P Harding J J Bomalaski J SGlassman D C Covington C M Brenner R Hollywood E Barba A et al(2017) A phase 11B trial of ADI-PEG 20 plus nab-paclitaxel and gemcitabine inpatients with advanced pancreatic adenocarcinoma Cancer 123 4556-4565
Luke J J LoRusso P Shapiro G I Krivoshik A Schuster R Yamazaki TArai Y Fakhoury A Dmuchowski C and Infante J R (2016) ASP9853 aninhibitor of inducible nitric oxide synthase dimerization in combination withdocetaxel preclinical investigation and a Phase I study in advanced solid tumorsCancer Chemother Pharmacol 77 549-558
Lundberg J O Weitzberg E and Gladwin M T (2008) The nitrate-nitrite-nitricoxide pathway in physiology and therapeutics Nat Rev Drug Discov 7 156-167
Maa M-C Chang M Y Chen Y-J Lin C-H Yu C J Yang Y L Li JChen P-R Tang C-H Lei H-Y et al (2008) Requirement of inducible nitric-oxide synthase in lipopolysaccharide-mediated Src induction and macrophagemigration J Biol Chem 283 31408-31416
MacMicking J Xie Q-W and Nathan C (1997) Nitric oxide and macrophagefunction Annu Rev Immunol 15 323-350
Magne J Huneau J F Delemasure S Rochette L Tome D andMariotti F(2009) Whole-body basal nitric oxide production is impaired in postprandialendothelial dysfunction in healthy rats Nitric Oxide 21 37-43
Marigo I Zilio S Desantis G Mlecnik B Agnellini A H R Ugel S SassoM S Qualls J E Kratochvill F Zanovello P et al (2016) T cell cancertherapy requires CD40-CD40L activation of tumor necrosis factor and induciblenitric-oxide-synthase-producing dendritic cells Cancer Cell 30 377-390
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
Marshall H E and Stamler J S (2001) Inhibition of NF-kappa B by S-nitrosylation Biochemistry 40 1688-1693
Martin E Davis K Bian K Lee Y C and Murad F (2000) Cellular signalingwith nitric oxide and cyclic guanosine monophosphate Semin Perinatol 24 2-6
Moreno-Morcillo M Grande-Garcia A Ruiz-Ramos A Del Cano-Ochoa FBoskovic J and Ramon-Maiques S (2017) Structural insight into the core ofCAD the multifunctional protein leading de novo pyrimidine biosynthesisStructure 25 912-923 e5
Mungrue I N Bredt D S Stewart D J and Husain M (2003) Frommoleculesto mammals whatrsquos NOS got to do with it Acta Physiol Scand 179 123-135
Murad F (2006) Shattuck Lecture Nitric oxide and cyclic GMP in cell signaling anddrug development N Engl J Med 355 2003-2011
Namin S M Nofallah S Joshi M S Kavallieratos K and Tsoukias N M(2013) Kinetic analysis of DAF-FM activation by NO toward calibration of a NO-sensitive fluorescent dye Nitric Oxide 28 39-46
Nath N Chattopadhyay M Rodes D B Nazarenko A Kodela R andKashfi K (2015) Nitric Oxide-releasing aspirin suppressesNF-kappaB signalingin estrogen receptor negative breast cancer cells in vitro and in vivoMolecules 2012481-12499
Ning S Bednarski M Oronsky B Scicinski J and Knox S J (2014) Novelnitric oxide generating compound glycidyl nitrate enhances the therapeuticefficacy of chemotherapy and radiotherapy Biochem Biophys Res Commun447 537-542
Paige J S and Jaffrey S R (2007) Pharmacologic manipulation of nitric oxidesignaling targeting NOS dimerization and protein-protein interactions Curr TopMed Chem 7 97-114
Pathi S S Jutooru I Chadalapaka G Sreevalsan S Anand S ThatcherG R and Safe S (2011) GT-094 a NO-NSAID inhibits colon cancer cell growthby activation of a reactive oxygen species-microRNA-27a ZBTB10-specificityprotein pathway Mol Cancer Res 9 195-202
Pershing N L K Yang C-F J Xu M and Counter C M (2016) Treatmentwith the nitric oxide synthase inhibitor L-NAME provides a survival advantage in amouse model of Kras mutation-positive non-small cell lung cancerOncotarget 742385-42392
Philippu A (2016) Nitric oxide a universal modulator of brain function Curr MedChem 23 2643-2652
Pipili-Synetos E Papageorgiou A Sakkoula E Sotiropoulou G Fotsis TKarakiulakis G and Maragoudakis M E (1995) Inhibition of angiogenesistumour growth and metastasis by the NO-releasing vasodilators isosorbidemononitrate and dinitrate Br J Pharmacol 116 1829-1834
Porasuphatana S Tsai P and Rosen G M (2003) The generation of freeradicals by nitric oxide synthase Comp Biochem Physiol C Toxicol Pharmacol134 281-289
Porcelli V Fiermonte G Longo A and Palmieri F (2014) The human geneSLC25A29 of solute carrier family 25 encodes a mitochondrial transporter ofbasic amino acids J Biol Chem 289 13374-13384
Predonzani A Cali B Agnellini A H and Molon B (2015) Spotlights onimmunological effects of reactive nitrogen species when inflammation says nitricoxide World J Exp Med 5 64-76
Puglisi M A Cenciarelli C Tesori V Cappellari M Martini M DiFrancesco A M Giorda E Carsetti R Ricci-Vitiani L and GasbarriniA (2015) High nitric oxide production secondary to inducible nitric oxidesynthase expression is essential for regulation of the tumour-initiating propertiesof colon cancer stem cells J Pathol 236 479-490
Rabender C S Alam A Sundaresan G Cardnell R J Yakovlev V AMukhopadhyay N D Graves P Zweit J and Mikkelsen R B (2015) Therole of nitric oxide synthase uncoupling in tumor progression Mol Cancer Res13 1034-1043
Rabinovich S Adler L Yizhak K Sarver A Silberman A Agron SStettner N Sun Q Brandis A Helbling D et al (2015) Diversion ofaspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis Nature527 379-383
Re A Aiello A Nanni S Grasselli A Benvenuti V Pantisano V StrigariL Colussi C Ciccone S Mazzetti A P et al (2011) Silencing of GSTP1 aprostate cancer prognostic gene by the estrogen receptor-beta and endothelialnitric oxide synthase complex Mol Endocrinol 25 2003-2016
Reddy K R K Dasari C Duscharla D Supriya B Ram N S SurekhaM V Kumar J M and Ummanni R (2018) Dimethylargininedimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostatecancer and its overexpression conveys tumor growth and angiogenesis bymetabolizing asymmetric dimethylarginine (ADMA) Angiogenesis 21 79-94
Rhee H J Kim E-J and Lee J K (2007) Physiological polyamines simpleprimordial stress molecules J Cell Mol Med 11 685-703
Ridnour L A Cheng R Y S Weiss J M Kaur S Soto-Pantoja D RBasudhar D Heinecke J L Stewart C A DeGraff W Sowers A L et al(2015) NOS inhibition modulates immune polarization and improves radiation-induced tumor growth delay Cancer Res 75 2788-2799
Rigas B andKashfi K (2004) Nitric-oxide-donating NSAIDs as agents for cancerprevention Trends Mol Med 10 324-330
Salimian Rizi B Caneba C Nowicka A Nabiyar A W Liu X Chen KKlopp A and Nagrath D (2015) Nitric oxide mediates metabolic coupling ofomentum-derived adipose stroma to ovarian and endometrial cancer cellsCancer Res 75 456-471
Segawa Y Oda Y Yamamoto H Uryu H Shiratsuchi H Hirakawa NTomita K Yamamoto T Oda S Yamada T et al (2008) Overexpression ofinducible nitric oxide synthase and accumulation of 8-OHdG in nasopharyngealcarcinoma Histopathology 52 213-223
Sektioglu I M Carretero R Bender N Bogdan C Garbi N Umansky VUmansky L Urban K von Knebel-Doberitz M Somasundaram V et al(2016) Macrophage-derived nitric oxide initiates T-cell diapedesis and tumorrejection Oncoimmunology 5 e1204506
Sestili P Ambrogini P Barbieri E Sartini S Fimognari C Calcabrini CDiaz A R Guescini M Polidori E Luchetti F et al (2016) New insightsinto the trophic and cytoprotective effects of creatine in in vitro and in vivo modelsof cell maturation Amino Acids 48 1897-1911
Seth D Hess D T Hausladen A Wang L Wang Y J and Stamler J S(2018) A multiplex enzymatic machinery for cellular protein S-nitrosylation MolCell 69 451-464 e6
Shan Y-S Hsu H-P Lai M-D Yen M-C Chen W-C Fang J-H Weng T-Y and Chen Y-L (2015) Argininosuccinate synthetase 1 suppression andarginine restriction inhibit cell migration in gastric cancer cell linesSci Rep 5 9783
Siemens D R Heaton J PW Adams M A Kawakami J andGraham C H(2009) Phase II study of nitric oxide donor for men with increasing prostate-specific antigen level after surgery or radiotherapy for prostate cancerUrology 74878-883
Singh R Avliyakulov N K Braga M Haykinson M J Martinez L SinghV Parveen M Chaudhuri G and Pervin S (2013) Proteomic identificationof mitochondrial targets of arginase in human breast cancer PLoS ONE 8e79242
Stettner N Rosen C Bernshtein B Gur-Cohen S Frug J Silberman ASarver A Carmel-Neiderman N N Eilam R Biton I et al (2018) Inductionof nitric-oxide metabolism in enterocytes alleviates colitis and inflammation-associated colon cancer Cell Rep 23 1962-1976
Syed N Langer J Janczar K Singh P Lo Nigro C Lattanzio L ColeyH M Hatzimichael E Bomalaski J Szlosarek P et al (2013) Epigeneticstatus of argininosuccinate synthetase and argininosuccinate lyase modulatesautophagy and cell death in glioblastoma Cell Death Dis 4 e458
Tachibana H Kakuta S Yagami K and Nagumo M (2000) Effects ofcytokines on the production of nitric oxide in a chondrogenic cell line establishedfrom human osteogenic sarcoma Oral Dis 6 303-309
Terzuoli E Finetti F Costanza F Giachetti A Ziche M and Donnini S(2017) Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells Nitric Oxide 66 17-29
Thomas D D Liu X Kantrow S P and Lancaster J RJr (2001) Thebiological lifetime of nitric oxide implications for the perivascular dynamics of NOand O2 Proc Natl Acad Sci USA 98 355-360
Thomas D D Espey M G Ridnour L A Hofseth L J Mancardi D HarrisC C and Wink D A (2004) Hypoxic inducible factor 1alpha extracellularsignal-regulated kinase and p53 are regulated by distinct thresholdconcentrations of nitric oxide Proc Natl Acad Sci USA 101 8894-8899
Toda N and Herman A G (2005) Gastrointestinal function regulation by nitrergicefferent nerves Pharmacol Rev 57 315-338
Tsai H-J Jiang S S Hung W-C Borthakur G Lin S-F Pemmaraju NJabbour E Bomalaski J S Chen Y-P Hsiao H-H et al (2017) A phase IIstudy of arginine deiminase (ADI-PEG20) in relapsedrefractory or poor-risk acutemyeloid leukemia patients Sci Rep 7 11253
Van de Wouwer M Couzinie C Serrano-Palero M Gonzalez-Fernandez OGalmes-Varela C Menendez-Antolı P Grau L and Villalobo A (2012)Activation of the BRCA1Chk1p53p21(Cip1Waf1) pathway by nitric oxide andcell cycle arrest in human neuroblastoma NB69 cells Nitric Oxide 26 182-191
Vasudevan D Hickok J R Bovee R C Pham V Mantell L L Bahroos NKanabar P Cao X-J Maienschein-Cline M Garcia B A et al (2015)Nitric oxide regulates gene expression in cancers by controlling histoneposttranslational modifications Cancer Res 75 5299-5308
Vasudevan D Bovee R C and Thomas D D (2016) Nitric oxide the newarchitect of epigenetic landscapes Nitric Oxide 59 54-62
Villalobo A (2006) Nitric oxide and cell proliferation FEBS J 273 2329-2344Wang C Gong G Sheh A Muthupalani S Bryant E M Puglisi D A
Holcombe H Conaway E A Parry N A P Bakthavatchalu V et al (2017)Interleukin-22 drives nitric oxide-dependent DNA damage and dysplasia in amurine model of colitis-associated cancer Mucosal Immunol 10 1504-1517
Watford M (1991) The urea cycle a two-compartment system Essays Biochem26 49-58
Weisser S B McLarren KW Kuroda E and Sly L M (2013) Generation andcharacterization of murine alternatively activated macrophages Methods MolBiol 946 225-239
Woodmansee A N and Imlay J A (2003) A mechanism by which nitric oxideaccelerates the rate of oxidative DNA damage in Escherichia coli Mol Microbiol49 11-22
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
Wu G Bazer F W Davis T A Kim S W Li P Marc Rhoads J CareySatterfield M Smith S B Spencer T E and Yin Y (2009)Arginine metabolism and nutrition in growth health and disease Amino Acids37 153-168
Wu L Li L Meng S Qi R Mao Z and Lin M (2013) Expression ofargininosuccinate synthetase in patients with hepatocellular carcinomaJ Gastroenterol Hepatol 28 365-368
Xie L and Gross S S (1997) Argininosuccinate synthetase overexpression invascular smooth muscle cells potentiates immunostimulant-induced NOproduction J Biol Chem 272 16624-16630
Yau T Cheng P N Chan P Chan W Chen L Yuen J Pang R Fan S Tand Poon R T (2013) A phase 1 dose-escalating study of pegylatedrecombinant human arginase 1 (Peg-rhArg1) in patients with advancedhepatocellular carcinoma Invest New Drugs 31 99-107
Zhang H Wang Q Gu J Yin L Liang S Wu L Xu H Zhao C and GuY (2018) Elevated mitochondrial SLC25A29 in cancer modulates metabolicstatus by increasing mitochondria-derived nitric oxide Oncogene 372545-2558
Zhao Y Vanhoutte P M and Leung S W S (2015) Vascular nitric oxidebeyond eNOS J Pharmacol Sci 129 83-94
![nitric oxide [15N]arginine-to-[15N]citrulline - pnas.org · period, in healthy subjects receiving an adequate arginine intake. Thisinvestigation establishes anexperimentalbasisfor](https://static.documents.pub/doc/80x56/5d402ba788c99377448bcf7f/nitric-oxide-15narginine-to-15ncitrulline-pnasorg-period-in-healthy.jpg)
