ROS and redox biology in cancer
- some preliminary efforts to synthesis based on selectedexemples
Credits to Elias Arner, Stig Linder, Rolf Kiessling, Rickard Holmdahl
Invitation:Please identify omissions and unclarities
Redox biology and cancer
Truism 1
Redox-regulatory proteins and ROS might have different and distinct functionsduring
- Cancer initiation- Primary tumor growth- Metastasis- Response to treatment
Redox biology and cancer
Truism 2
Redox-regulatory proteins and ROS might have different and distinct functionsin malignant cells and in cells of the tumor microenvironment
The tumor is an organ composed of multiple cell types reciprocal paracrine signaling govern tumor behavior
inflammatory cells
reviewed in Östman Augsten, Curr Op Gen Dev, 2009: Pietras Östman, Exp Cell Res, 2010; Östman, Nat Med, 2012; Östman, Sem Can Biol, 2014; Östman, ADDR, 2018; Corvigno, Östman, Trends in Cancer, 2018
ROS and cancer biology
Two selected aspects:
1. Oxidative stress and ROS in malignant cells during tumor initiation and progression
2. ROS-mediated suppression of immune surveillance
ROS/oxidative stressin malignant cells
ROS drivescancer formation
through mutagenic effects
Oxidative stressin cancer requires
an active anti-oxidantdefence
ROS/oxidative stressin malignant cells
ROS drivescancer formation
through mutagenic effects
Radiation
Mutagens
UV
Chemotherapy induced secondary cancer
Oxidative stressin cancer requires
an active anti-oxidantdefence
hypoxia/altered metabolism/oncogene activation
mitochondrial ROS/NOX-derived ROS
ROS drivescancer formation
through mutagenic effects
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancer drugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
1. KEAP loss of function mutation/Nrf2 gain of function mutation in human cancer
2. Pro-migratory and metastatic effects of anti-oxidants
3. Novel candidate cancer drugs targeting the ”oxidative stress vulnerability” of malignant cells
4. Low-expression/loss of Nrf2 is associated with increased lung cancer risk
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancerdrugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
ROS drivescancer formation
through mutagenic effects
Yamamoto, Physiol Rev, 2018
The KEAP1-NRF2 SYSTEM
Loss-of-function mutations of KEAP and gain-of-function mutation of Nrf2have been detected in human cancer
Shibata, PNAS, 2008 Singh, PLoS Med, 2006
GOF Nrf2 mutations in 10% of lung cancer LOF KEAP mutations in 20% of lung cancer
Anti-oxidant Nrf2 system is cancer-driving
alterations in KEAPinteraction sites
Overall mutations; high frequency of stop codons
Mutation profiles from human tumors support the notion thatKEAP variants are loss-of-function and NRF2 variants are gain-of function
Yamamoto, Physiol Rev, 2018
Solis, ClinCanRes, 2010
Analyses of clinical samples reveal poor prognosis-associations of Nrf2 in malignant cells
Anti-oxidant Nrf2 system is cancer-driving
Okano, PLoS One, 2013
Low expressing Nrf2 variant is good prognosis marker in lung cancer
DeNicola, Nature, 2011
Nrf2 is activated in ras-dependent mouse pancreas cancer modeland support tumor growth
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancer drugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
ROS drivescancer formation
through mutagenic effects
1. KEAP loss of function mutation/Nrf2 gain of function mutation in human cancer
2. Pro-migratory and metastatic effects of anti-oxidants
3. Novel candidate cancer drugs targeting the ”oxidative stress” vulnerability of malignant cells
4. Low-expression/loss of Nrf2 is associated with increased lung cancer risk
Sayin, Sci Transl Med, 2014Le Gal, Sci Transl Med, 2015
Enhanced growth, migration and metastasis after anti-oxidanttreatment of mouse melanoma
migration metastasis
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancer drugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
ROS drivescancer formation
through mutagenic effects
1. KEAP loss of function mutation/Nrf2 gain of function mutation in human cancer
2. Pro-migratory and metastatic effects of anti-oxidants
3. Novel candidate cancer drugs targeting the ”oxidative stress” vulnerability of malignant cells
4. Low-expression/loss of Nrf2 is associated with increased lung cancer risk
Schoenfeld, Cancer Cell, 2017
Schoenfeld, Cancer Cell, 2017
Feasibility and preliminary efficacy of high dose ascorbate(pro-oxidant) therapy in GBM used with standard chemo and radiation
Ngo, Nat Rev Cancer, 2019
Table listing20plus clinical trials
Stafford, SciTranslMed, 2018
Cancer cell specific killing of novel TrxR1inhibitor
Halufuginon; a novel Nrf2 inhibitor with cancer drug potential
Tushida, FRBM, 2017
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancerdrugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
ROS drivescancer formation
through mutagenic effects
1. KEAP loss of function mutation/Nrf2 gain of function mutation in human cancer
2. Pro-migratory and metastatic effects of anti-oxidants
3. Novel candidate cancer drugs targeting the ”oxidative stress” vulnerability of malignant cells
4. Low-expression/loss of Nrf2 is associated with increased lung cancer risk
Suzuki, MCB, 2013
An Nrf2 variant, associated with reduced Nrf2 expression, confers highrisk for lung cancer development
Nrf2-/- mice are hyper-sensitive to chemically induced cancer
Chemically induced colon cancer
Iida, Can Res, 2004
Khor, Can Prev Res, 2008
Ramos-Gomez, PNAS,2001
Satoh, Can Res, 2013
Increased number of cancer lesions, but reduced metastasis, afterChemical lung cancer induction in Nrf2-/- mice
ROS/oxidative stressin malignant cells
Oxidative stressin cancer requires
an active anti-oxidantdefence
Pro-oxidants are cancerdrugs
Anti-oxidants are cancer-driving
Pro-oxidants are cancer promoting
Anti-oxidants are cancer-protective
ROS drivescancer formation
through mutagenic effects
1. KEAP loss of function mutation/Nrf2 gain of function mutation in human cancer
2. Pro-migratory and metastatic effects of anti-oxidants
3. Novel candidate cancer drugs targeting the ”oxidative stress” vulnerability of malignant cells
4. Low-expression/loss of Nrf2 is associated with increased lung cancer risk
ROS and cancer biology
Two selected aspects:
1. Oxidative stress and ROS in malignant cells during tumor initiation and progression
2. ROS-mediated suppression of immune surveillance
ROS/oxidative stressand tumor immune surveillance
-overall concept
Anti-oxidants can be anti-tumoral through activation of immune surveillance-activation CD8 T-cells-inactivation of Tregs
-inactivation of MDSCs-activation of dendritic cells
1. Human/mouse studies implying Ncf1 (Nox2) as immune suppressive
2. ROS-mediated MDSC-induced immune supression- inactivation/killing of T-cells
- -enrichment of Tregs
3. Nrf2-mediated immune regulation
4. Therapeutic opportunities
Nox 2 down ROS down T-cell activity up auto-immune disese
NOX2-derived ROS act as immune suppresant
Nox 2 down ROS down T-cell activity up auto-immune disese
NOX2-derived ROS act as immune suppresant
Host-depletion of NOX 2 decreases tumor growth
Data still pending for effects of cell-type-specific Ncf1 deletion
Kelkka, PLoS One, 2013
Interpretation:Host Nox2 deficiency – improved immune surveillance
Host-depletion of NOX 2 decreases metastasis formation
Metastasis formation Immune cell infiltration
van der Weyden, J Path, 2018
Interpretation:Host Nox2 deficiency – improved immune surveillance
1. Human/mouse genetics implying Ncf1 (Nox2) as tumor immune suppressive
2. ROS-mediated MDSC-induced tumor immune suppression
3. Nrf2-mediated immune regulation
4. Therapeutic opportunities
ROS/oxidative stressand tumor immune surveillance
-overall concept
Anti-oxidants can be anti-tumoral through activation of immune surveillance-activation CD8 T-cells-activations of NK cells
A
Mao, JIM. 2014
MDSC-derived ROS suppress T-cell and NK-cell activityNOTE paracrine ROS signaling
T cell compartment
CD4+ CD8+
TEM
TEM
TCM
TCM
TEMRA
NaïveNaïve
Sen
siti
vity
to
H2O
2-i
nd
uce
d c
ell
de
ath
NK cell compartment
CD56bright CD56dim
The T- and NK cell subsets which are the most efficient in
eliminating tumor cells are the most sensitive ones to oxidative
stress.
Differential sensitivity to oxidative stress among lymphocyte subsets.
Effector T-cells are hyper-sensitive toROS-induced cell death
- implications: make better effector T-cells by improving their anti-oxidant defence
Takahashi, J. Imm, 2005
1. Human/mouse genetics implying Ncf1 (Nox2) as tumor immune suppressive
2. ROS-mediated MDSC-induced tumor immune suppression
3. Nrf2-mediated immune regulation
4. Therapeutic opportunities
ROS/oxidative stressand tumor immune surveillance
-overall concept
Anti-oxidants can be anti-tumoral through activation of immune surveillance-activation CD8 T-cells-activation of NK cells
Loss of host Nrf2 activity is associated withincreased metastasis outgrowth
Gain of host Nrf2/loss of KEAPactivity is associated withreduced metastasis outgrowth
Satoh, Carcinogenesis, 2010
Possible interpretations
Low Nrf2:”weak/attenuated/few” T-cells;
increased metastasis
high Nrf2:”strong/active/many” T-cells;
decreased metastasis
1. Human/mouse genetics implying Ncf1 (Nox2) as tumor immune suppressive
2. ROS-mediated MDSC-induced tumor immune suppression
3. Nrf2-mediated immune regulation
4. Therapeutic opportunities
ROS/oxidative stressand tumor immune surveillance
-overall concept
Anti-oxidants can be anti-tumoral through activation of immune surveillance-activation CD8 T-cells-activation of NK cells
Ando, J. Imm, 2008
ROS-tolerance of T-cells can be increased bycatalase over-expression
A positve phase III trial possibly involvingprotection from ROS induced killing of NK cells
Brune, Blood, 2006
ROS/oxidative stressand tumor immune surveillance
-overall concept
Anti-oxidants can be anti-tumoral through activation of immune surveillance-activation CD8 T-cells-activation of NK cells
ROS/redox/Nrf2and CAFs??????
ROS/Redox/Nrf2and tumor vasculature??????
Teaser for next course
Cell-type specific redox profiling of a well-annotatedlung cancer collection
Background:
oxidative stress has been suggested to have different functional impact on different cell types
prognostic impact of cell-type-specific oxidative stress remain unknown
relationships between cell type specific oxidative stress and driver mutations and immune surveillance remain unnown
Aim of study:
to explore the prognostic significance in lung cancer of oxidative stress in different marker-defined tumor cell types
to perform explorative correlative analyses that can suggest biological mechanisms that drive the prognosticallysignificant cell-type-specific oxidative stress
Immune cellcomposition
IF/ Marker panel
Cell types
Pan-cytokeratin tumor cells
Alpha-SMA fibroblasts, perivascular cells
CD68 macrophages
CD3 T-cells
Oxidative stress
Nrf2
TrxR1
”Redox metrics”
Tumor epithelial cells fraction ox stress pos total tumor areaarea density ox stress pos juxta-stromal tumor epithelial area
tumor epithelial area 1 x2x3=6
CD68 cells/ fraction ox stress pos total tumor areaCD3 cells area density ox stress pos stroma area
juxta epithelial stroma areaepithelial areajuxta-stromal epithelial area 2x2x5=20
ASMA + cells fraction ox stress pos total tumor areaarea density ox stress pos stroma area
juxta-epithelial area 1x2x3=6
32 metrics
( for either Nrf2 or TrxR1)
Analyses plan
32 Nrf2 metrics
32 TrxR1 metricsvs overall survival (prognosis)
smoking/etiologydriver mutationsimmune phenotype
ROS/redox/Nrf2and CAFs??????
ROS/Redox/Nrf2and tumor vasculature??????
Credits to Elias Arner, Stig Linder, Rolf Kiessling, Rickard Holmdahl
Course organizers and participants
