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Dept. of Pediatrics and Adolescent MedicineInflammation and Metabolic Disease
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Inflammation and Metabolic Disease
Maximilian Zeyda, PhD
Dept. of Pediatrics and Adolescent Medicine
Newborn Screening and Metabolic Lab
Basic lecture (BVO) „MolSignTrans“ 2018
2Inflammation and Metabolic DiseaseM Zeyda
small molecular alteration -> huge consequences
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Outline
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• Interrelation metabolism/inflammation on signaling level
• Insulin resistance and implications
• Obesity-induced adipose tissue inflammation as a cause of
metabolic disease
• Mechanisms underlying obesity-induced inflammation
Osteopontin as an example for a molecular player in obesity-
associated inflammation and for a potential treatment target
Inflammation and Metabolic DiseaseM Zeyda
How does inflammation interrelate with insulin-
dependent metabolism?
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Kumar M., et al. Protein Cell. 2012 3(10):726‐38
Insulin signaling
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Kwon, Frontiers in Immunology 2013
Inflammatory signaling blocks insulin signaling ‐> IR
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Also TLR signaling blocks insulin signaling
OPN
Osteopontin (OPN; gene Spp1), is a multifunctional proteinexpressed in osteoclasts, hepatocytes, smooth muscle cells, endothelial cells, epithelial cells, and activated T cells and macrophages
OPN was classified as a T helper type 1 (Th1) cytokine that is involved in chronic inflammatory disorders as well as in tumor biology
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OPN signaling blocks insulin signaling
Osteopontin (OPN) actssimilar to TNF on insulin‐stimulated glucose uptake of human adipocytes
Zeyda et al. Endocrinology 2011
..via integrin signaling
From Humpgries et al., Journal of Cell Science 2006 119: 3901‐3903
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Wenjun Guo & Filippo G. Giancotti, Nature Reviews Molecular Cell Biology volume 5, pages 816–826 (2004
• Chronically inflammatory signaling‐> chronically blocked insulin signaling‐> insulin resistance‐> the road to type 2 diabetes
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…and further complications
Does block of insulin sensitivity by inflammation make sense (in a biological/physiological view)?
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Warburg Metabolism (1924)
Heiden et al. Science 2009
Also effector T cells use aerobic glycolysis
Science (New York, N.Y.). 2013 342(6155):1242454
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M1/M2 polarization affects metabolism: M1 need glucose (and oxygen)
Vats et al., Cell Metabolism 2006 O’Neill and Hardie Nature 2013
Conclusions (so far)
• Inflammation blocks insulin signaling, which may be of biological advantage (in acute host defense)
• Chronically blocked insulin signaling means insulin resistance
• Insulin resistance is the basis for type 2 diabetes and a plethora of diseases
• What´s the problem as long we don´t suffer from a chronic inflammatory disease?
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Obesity(is a chronic inflammatory disease)
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Why bother: Obesity‐associated diseases
• Diabetes type 2
• Cardiovascular disease
• Dyslipidemia
• Hypertension
• Cancer
• ...
Metabolic SyndromeCardio-Metabolic Risk
• strong association with several
specific causes of mortality *
• Increased incidence in obesity‐
asociated disorders like type 2
diabetes, hypertension,
coronary heart disease, and
cholelithiasis** …
* Whitlock et al. – Lancet, 2009** Willett et al. – NEJM, 1999
Obesity (BMI)
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Diabetes Prevalence Worldwide
International Diabetes Federation. The IDF Diabetes Atlas. Fourth Edition, 2011
366 Mio individuals worldwide8,3% of adults
Don´t underestimate thehealth problem!
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Cost explosion by obesity-associateddiseases, US data
SL Wang, Lancet 2011;378:815-25extrapolated based on historical trend
Why are these diseases associated with obesity?
Key is the adipose tissue
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Adipose tissue is the organ of fat storage
Nutrient overload ‐> adipose tissue insulin resistance ‐> elevated free fatty acids ‐> ectopic fat accumulation
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“Insulin Sensitivity: Visceral or SC Fat – That’s the Question!”, N Klöting, Am J Physiol Endocrinol Metab 2010;299:E506‐15
The risk depends on the distribution of body fat
sc – stable energy store intrabdominal – more mobile
More than fat storage: AT is an endocrine organ
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Nature Immunology 13, 707–712 (2012)
Obesity is an inflammatory condition resulting from inflamed adipose tissue
• Clinically recognized for a long time (inflammatory parameters upregulated in obese, correlation of inflammation with Type 2 Diabetes)
• Hotamisligil, GS, Shargill, NS, Spiegelman, BM. Adipose expression of tumor necrosis factor‐alpha: direct role in obesity‐linked insulin resistance. Science. 1993. 259:87‐91.
• Xu, J Clin Inv 2003, Weisberg, J Clin Inv 2003:
Macrophages accumulate in adipose tissue of obese mice
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Stuart P. Weisberg, …, Anthony W. Ferrante Jr.J Clin Invest. 2003; 112(12):1796–1808
OPN upregulation in obesity
0
1000
2000
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4000
5000
6000
mRNA expression in
AT
(% of LF)
HFLF isotype control
A
C
OPN
**
0
50
100
150
200
LF HF
**
OPN mRNA expression
in liver (% of LF)
F4/80 TNF Mcp-1
*******
B0
2000
4000
6000
8000
10000
12000
mRNA expression in
AT
(% of db/+)
A
OPN
*
db/+ db/db0
n.s.
50
100
150
200
OPN mRNA expression
in liver (% of db/+)
CD68 TNF Mcp-1
**** **
B
0
500
1000
1500
2000
2500
lean obese
OPN mRNA expression
(% of lean
omen
tum)
omentumlean obese
subcutis
**
***
diet‐induced obese mice genetically obese mice humans
Kiefer et al. Endocrinology 2008
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Insulin Sensitivity Correlates with Inflammation-1 and Adiponectin
N Klöting, Am J Physiol Endocrinol Metab 2010;299:E506-15
Biochemical Journal 2010 430, e1-e4 - Marie-Soleil Gauthier and Neil B. Ruderman
Inflamed adipose tissue affects the whole organism
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Kiefer et al. Diabetologia 2011
OPN deficiency improves obesity‐associatedadipose tissue inflammation, hepatic steatosis, and systemic and hepatic insulin resistance
Koliaki et al. Metabolism. 2018 Nov 3. pii: S0026‐0495(18)30229‐4.
Example: Obesity and cardiovascular diseaseIt´s about adipose tissue (immune) function
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OK, inflammation (and within inflammation, OPN may be important) is the key, but still: how does obesity
cause inflammation?
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Endotoxemia may play a role
Metabolic endotoxaemia: is it more than just a gut feeling?.Piya, Milan; Harte, Alison; McTernan, PhilipCurrent Opinion in Lipidology. 24(1):78‐85, February 2013.
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..together with various factors affected by nutrition and the gut microbiome
Zhang et al. Science Reports 2017
White adipose tissue (WAT) macrophages localize to crown‐like structures (CLS) around individual adipocytes,
their frequency increases with obesity.
Cinti S et al. J. Lipid Res. 2005;46:2347-2355
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CLS form exclusively at sites of adipocyte death and scavenge the residual adipocyte lipid droplet.
Cinti S et al. J. Lipid Res. 2005;46:2347-2355
Zeyda et al. Int J Obes (Lond). 2010;34(12):1684‐94
Obesity causes adipocyte hypertrophy that causes ROS production and ER stress in adipocytes ‐> inflammatory response
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Russo L, Lumeng CN. Properties and functions of adipose tissue macrophages in obesity. Immunology. 2018 155(4):407-417
Han and Levings. Immune regulation in obesity‐associated adipose inflammation. Journal of immunology. 2013 191(2):527‐32Winer and Winer, Immun and Cell Biol 2012
innate lymphos (?)NKT cells
Obesity induces a shift from M2 to M1 (and all kind of immune cells are involved)
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…affecting, e.g., the liver (NAFLD)
Nutrients 2017, 9(4), 387
..and, of course, the inflammasome is involved
Nature Immunology 13, 707–712 (2012)
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..and, and, and…
Free DNAObesity‐induced DNA released from adipocytes stimulates chronic adipose tissue inflammation and insulin resistanceSachiko Nishimoto, et al. Science Advances 2016; 2(3):e1501332
miRNAmiR‐146a‐mediated suppression of the inflammatory response in human adipocytes.Roos J, et al. Sci Rep. 2016; 6:38339
Browning, beigingInflammation of brown/beige adipose tissues in obesity and metabolic disease
F. Villarroya, et al., J Intern Med. 2018; 284(5):492‐504.
miRNA and browningObesity‐Associated miR‐199a/214 Cluster Inhibits Adipose Browning via PRDM16–PGC‐1α Transcriptional NetworkLinyun He, et al. Diabetes 2018; 67(12): 2585‐2600.
Secreted factors that induce browning
Nature Medicine 19, 1252–1263 (2013)
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Kwon, Frontiers in Immunology 2013
OPN
M. Tardelli, et al. Mol Metab. 2016
Apoptosis Proliferation
Human monocytes are protected from apoptosisand proliferate in presence of OPN
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Also fully differentiated human macrophagesproliferate in response to OPN
M. Tardelli, et al. Mol Metab. 2016
M. Tardelli, et al. Mol Metab. 2016
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8 wks 12 wks
Macrophages in adipose tissue proliferate less in OPN‐deficient mice
BMDM proliferation is independentof intrinsic OPN
M. Tardelli, et al. Mol Metab. 2016
adipocyte
hypertroph adipocyte
resident Mactivated M
vasculaturemonocyte
Lean
regular turnover
Obese
insulin resistanceIR
* necrotic adipocyte
OPNpolarizationto phagocytic, protective phenotype
proteasecleavage
*
OPN
recruitment ofmonocytesby multiple factors, enhanced by OPN
OPN
IRSurvivalproliferation
IR
Summary OPN
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Current Strategies to Reduce Cardio‐Metabolic Risk
• Life style modification (diet, exercise, no smoking)
– limited effectiveness in clinical practice
• Pharmacologie weight reduction
– Appetite suppressants withdrawn
– Fat resorption inhibitor of limited effect
• Bariatric surgery
– effective, limited capacities, malnutrition/deficiencies
• Pharmacologic prevention of type 2 diabetes– sulfonylureas and/or metformin with ancillary use of acarbose and
thiazolidinediones
– Reduction of cardiovascular events??
• Drugs to reduce cardiovascular risk
– Lipid‐lowering drugs (statins); antihypertensive drugs
Immunotherapy (?)
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Immunotherapy against proteins naturallyproduced in the body
Immunotherapy against OPN?
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0 30 60 90 120
Time (min)
Blo
od g
luco
se (
mg/
dl)
**
#
80
90
100
110
120
130
140
Are
a un
der
the
curv
e
150
Control Anti‐OPN
*
A B
Kiefer et al., Diabetes 2010
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The central region 158GRGDSVVYGLR168
Highly conserved GRGDS integrin binding site commonly found in matrix proteins followed by thrombin andMMP cleavage sites YG166/LR168/SK
Protease cleavage unmasks the “cryptic” SVVYG(LR) sequence, a binding site for integrins prevalent on leukocytes not accessible in the full length protein
These characteristics together with the fact that MMP and thrombin activity is an aspect of inflammation offers interesting possibilities to target inflammation‐specific functions of cleaved OPN
Yamaguchi et al. J Biol Chem 2013
adapted from: Rittling et al., Expert Reviews in Molecular Medicine 2011
thrombinMMP
OPN cOPN
Increased abundance of cleaved OPN and presence of free MMP cleavage site in obese human AT.
Leitner et al. Obesity 23: 779‐785; 2015
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Protease cleavage increases OPN‐induced adipocyte inflammation and insulin resistance
Leitner et al. Obesity Volume 23, Issue 4, pages 779‐785, 16 MAR 2015
HEK cell adhesion is enhanced to truncated forms of OPN corresponding to thrombin
and MMP cleavage
Jürets et al., PlosOne 2016
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mAb 21‐5 blocks adhesion to both cleaved OPN forms, mAb 9‐3 is specific for mOPN
Binding of mAbs to immobilized OPN forms
mAbs included in adhesion assay
Jürets et al., PlosOne 2016
Immunological blockade of adipocyte inflammation caused by increased matrix metalloproteinase‐cleaved osteopontin
Leitner et al., ObesityVolume 23, Issue 4, pages 779‐785, 16 MAR 2015 DOI: 10.1002/oby.21024http://onlinelibrary.wiley.com/doi/10.1002/oby.21024/full#oby21024‐fig‐0006
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Jürets et al., PlosOne 2016
Sera from mice immunized with respective peptides can selectively block HEK adhesion to
truncated OPN forms
Inflammation, Metabolic Disease, OPN, immunotherapy - Summary
• Obesity causes a chronic inflammation originating in adipose tissue due to complex mechanisms not entirely understood
• Inflammation blocks insulin signaling
• Chronically blocked insulin signaling means insulin resistance, which is the basis for type 2 diabetes and a plethora of diseases
• Therefore, obesity-associated inflammation is a basis for cardiometabolic risk and should be targeted
• Immunotherapy could be an innovative approach and affordable (active IT) approach to neutralize inflammatory factors
• OPN with it´s cryptic (neo-)epitopes represents an interesting target
• Several studies were performed with promising results but active vaccination did not succeed so far [N. Grün et al., Immunol. Letters 2016]. Species differences may be the reason, therefore, vaccinations in a humanized mouse
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murine Spp1 signal peptide
Mouse genomic locus
murine Spp1 untranslated region
murine Spp1 coding exon
1 2 3 4 5 6 7
mouse genomic region
Targeted allele
1 2
NeoR
3 4 5 6 7
PuroR
human genomic region
FRT site F3 site
human SPP1 coding exonHumanized allele
1 2 3 4 5 6 7
A
B
The humanized OPN mouse
N. Grün, N. Grün et al., Transl Res. 2016
72Inflammation and Metabolic DiseaseM Zeyda
Thank you for your attention!!
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