Adipose tissue, innate immunity and inflammation – a nutrigenomics perspective of the metabolic syndrome Lecture in Scheveningen 2 Sept 2011

1. Adipose tissue, innate immunity and inflammation – a nutrigenomics perspective of the metabolic syndrome http://twitter.com/nutrigenomics Michael MüllerNetherlands Nutrigenomics Centre & Nutrition, Metabolism and Genomics GroupDivision of Human Nutrition, Wageningen University

2. I will talk about Our challenges: What is healthy What is Nutrigenomics? The metabolic syndrome The deadly sins Good fat / bad fat (tissue) Modern nutritional science & early biomarkers NASH & the role of the adipose tissue Dietary saturated fat can induce pro-inflammatory responses We have different phenotypes: Personalized health Saturated fat can be “killing” (in transgenic mice) Summary & recommendations

3. What do we know about the health network?

4. Our scientific challenge: What's healthy?

5. We are what we eat

6. Our “paleolithic” genes + modern diets Paleolithic era Modern Times 1.200.000 Generations between feast en famine 2-3 Generations in energy abundance % Energy % Energy 100 100 Grain Milk/-products Isolated Carbohydrates Isolated Fat/OilAlcohol Low-fat meatChicken Eggs Fish 50 Meat Chicken Fish 50 Fruit Vegetables (carrots) Nuts Honey Fruit Vegetables Beans 0 0

7. Nutrigenomics Quantification of the nutritional genotype-phenotype Lifestyle Nutrition Environment

8. Adipocytes at the crossroads of energy homeostasis

9. What is the metabolic syndrome? The metabolic syndrome is characterized by a group of metabolic risk factors in one person: Central obesity (excessive fat tissue in and around the abdomen) Atherogenic dyslipidemia (blood fat disorders — mainly high triglycerides and low HDL cholesterol — that foster plaque buildups in artery walls) Raised blood pressure (130/85 mm Hg or higher) Insulin resistance or glucose intolerance (the body can’t properly use insulin or blood sugar) Prothrombotic state (e.g., high fibrinogen or plasminogen activator inhibitor in the blood) Proinflammatory state (e.g., elevated high-sensitivity C-reactive protein in the blood)

10. The 7 deadly sins (gluttony & sloth)

11. Proposed pathway to the metabolic syndrome

12. Metabobolic homeostasis & syndrome

13. Normal Type 2 Diabetes Visceral Fat Distribution:Normal vs Type 2 Diabetes

15. We need a new nutritional science Insulin ± oral agents Oral combination Oral monotherapy Diet & exercise Complex Disease 100 Different & similar targets 80 Pharma 60 DISEASE STATE (%) 40 Nutrition 20 0 TIME (months/years) HomeostasisHealth L. Afman & M. Müller J Am DietAssoc. 2006;106:569-576.

16. Late biomarkersof disease Earlybiomarkersof disease Onset of disease Biomarkers of earlydiseasestate Single marker vsmultimarker profiles Disease Pharma Nutrition Early biomarkers in human nutrition research healthy

17. Organ-specific gene expression signatures of the early phase (metabolic stress) & the late phase of metabolic syndrome 1 2 3 4 10 16 Weeks WAT 1 2 3 4 10 16 Weeks Muscle 1 2 3 4 10 16 Weeks Liver 1 2 3 4 10 16 Weeks Intestine Healthy Unhealthy Healthy Unhealthy Healthy Unhealthy Healthy Unhealthy

19. Metabolism & Inflammation

21. Different stages in NAFLD progression:

22. Molecular events involved in NASH pathogenesis:

23. Role of PPARa (Endocrinology 2008 & Hepatology 2010)

24. Role Kupffer cells (Hepatology 2010)

25. Role of macrophages in lipid metabolism (JBC 2008; Cell Metabolism 2010)hepatic steatosis steatohepatitis (NASH) & fibrosiscirrhosis

26. Study: Interaction between WAT and liver tissue essential for NASH/NAFLD in C57Bl/6 mice Objective: Nonalcoholic fatty liver disease (NAFLD) is strongly linked to obesity and diabetes, suggesting an important role of adipose tissue in the pathogenesis of NAFLD. Here we aimed to investigate the interaction between adipose tissue and liver in NAFLD, and identify potential early plasma markers that predict NASH.

29. High fat diet-induced obesity 0 2 4 8 12 16 20 HFL LFL HFH LFH 25 20 * * 15 ** BW gain (g) * 10 * * * * 5 0 weeks under diet intervention Liver TG content Hepatomegaly ALT plasma activity 200 10 100 *** *** ** 160 8 80 ** 120 6 60 * Ratio LW/BW (%) mg TG/g liver ALT activity (UI) 80 4 40 * * 40 2 20 0 0 0 LFL LFH HFL HFH

30. A subpopulation of mice fed HFD develops NASH

31. Immunohistochemicalstaining confirms enhanced liver inflammation and early fibrosis in HFH mice Macrophage CD68 Collagen Stellate cell GFAP

32. Upregulation of inflammatory and fibrotic gene expression in HFH responder mice

33. Adipose dysfunction in HFH mice

34. Change in adipose gene expression indicate adipose tissue dysfunction

35. Plasma proteins as early predictive biomarker for NASH in C57Bl/6 mice

36. Plasma proteins as early predictive biomarker for NASH in C57Bl/6 mice Multivariate analysis of association of protein plasma concentrations with final liver triglyceride content

37. Conclusions Our data support the existence of a tight relationship between adipose tissue dysfunction and NASH pathogenesis. It points to several novel potential predictive biomarkers for NASH. Duval C, Thissen U, Keshtkar S, Accart B, Stienstra R, Boekschoten MV, Roskams T, Kersten S, Müller M. Adipose tissue dysfunction signals progression of hepatic steatosis towards nonalcoholic steatohepatitis in C57BL/6 mice. Diabetes. 2010;59:3181-91.

39. Human nutrigenomics study Dietary fat and inflammation in adipose tissue Change in diet composition ? Van Dijk et al. AJCN 2009 de Luca, C and Olefsky JM, Nature Medicine 12, 41 - 42 (2006)

41. Adipose tissue biopsy

43. Adipose tissue biopsy

44. Blood samplingVan Dijk et al. AJCN 2009

45. ‘Obese-linked’ pro-inflammatory gene expression profile by SFAs MUFA diet SFA diet The SFA-rich diet: Induces a pro-inflammatory obese-linked gene expression profile Decreases expression and plasma level of the anti-inflammatory cytokine adiponectin “Personal Transcriptomes” Van Dijk et al. AJCN 2009

46. Humanstudy:Plasma Protein Profiling Reveals Protein Clusters Related to BMI and Insulin Levels in Middle-Aged Overweight Subjects AIM Associate plasma protein profiles with BMI Identifypotential marker profile of earlydisease state . PLoS One. 2010 Dec 23;5(12):e14422

47. Measurements RulesBasedMedicine (Austin, USA) Multiplex immunoassay In total 124 proteinsmeasured Involved in diseases, inflammation, endothelialfunction and metabolism . PLoS One. 2010 Dec 23;5(12):e14422

48. We are different: improved phenotyping necessary to reveal phenotype clusters . PLoS One. 2010 Dec 23;5(12):e14422

49. Conclusion We identified clusters of plasma proteins associated with BMI and insulin in a healthy population. These clusters included earlier identified biomarkers for obesity-related disease as well as potential new biomarkers. These plasma protein clusters could have potential applications for improved phenotypic characterization of volunteers in nutritional intervention studies or as biomarkers in the early detection in obesity-linked disease development and progression. van DijkSJ, Feskens EJM, Heidema AG, Bos MB, van de Rest O, Geleijnse JM, de Groot CPGM, Müller M, Afman LA. Plasma Protein Profiling Reveals Protein Clusters Related to BMI and Insulin Levels in Middle-Aged Overweight Subjects. PLoS One. 2010 Dec 23;5(12):e14422

50. Chylomicron CE/TG Angptl4 LPL CE/TG FFA Chylomicron remnant

51. Angptl4- mice on HFD become very ill Lichtenstein et al. Cell Metab. 2010

52. Inflammatory response independent of microbiota Lichtenstein et al. Cell Metab. 2010

53. No effect of medium chain or PUFA TGs Lichtenstein et al. Cell Metab. 2010

54. Massive enlargement of mesenteric lymph nodes in Angptl4-/- mice fed HFD

55. Angptl4 inhibits lipolysis and subsequent foam cell formation

57. MLN-resident macrophages are protected from the pro-inflammatory effect of saturated fatty acids via expression of Angptl4, which is strongly induced by chyle and fatty acids and which via inhibition of LPL prevents lipolysis of chylomicron-TG.

58. In the absence of this protective mechanism, feeding a diet rich in saturated fat rapidly leads to enhanced lipid uptake into MLN-resident macrophages, triggering foam cell formation and a massive inflammatory response.Lichtenstein et al. Cell Metab. 2010

59. Is bariatric surgery the only solution?

60. Pharma is not the (only) solution:Eat foods rich in challenging food bioactives Drugs A B C PPARg PPARb PPARa Receptor C3 C2 C1 Fatty acids F C6 C5 C4 Multiple targets

61. Summary You are what you eat => during life all events will leave their (epigenetic) traces on our genome, some are irreversible => ageing Disease phenotypes such as obesity, metabolic syndrome, diabetes are largely related to our “gluttony / sloth” lifestyles and modern convenient (“fast”) foods => unhealthy ageing. NASH is the liver phenotype of the metabolic syndrome and appears early in the progression towards diabetes or CVD. There is a tight relationship between adipose tissue dysfunction and NASH pathogenesis. Chronic overconsumption of saturated fat or lipogenic precursors (starch, sugars) induces non-resolving low-grade pro-inflammatory state largely caused by the innate immune system. => choose the right lifestyle & food pattern (diverse & anti-inflammatory), eat less & exercise more (at least for 30 minutes/day).

62. 2 Meals a day, work as long as possible & embrace challenge Walter Breuning (1896 - 2011)

63. Sander KerstenLinda SandersonNatasha Georgiadi Mark BouwensLydia Afman Guido Hooiveld Meike Bunger Philip de Groot Mark Boekschoten Nicole de Wit Mohammad Ohid Ullah Christian Trautwein Folkert Kuipers Ben van Ommen + many more