Nutrigenomics of FAT: What is “good” or “bad” for human health?

Michael MüllerNetherlands Nutrigenomics Centre

& Nutrition, Metabolism and Genomics GroupDivision of Human Nutrition, Wageningen University

2 Meals a day, work as long as possible & embrace challenge

Walter Breuning (1896 - 2011)

We have a tsunami of health problems

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Low-fat meatChickenEggsFish

FruitVegetables (carrots)NutsHoney

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GrainMilk/-productsIsolated CarbohydratesIsolated Fat/OilAlcohol

1.200.000 Generations between feast en famine

Paleolithic era

2-3 Generations in energy abundance

Modern Times

Our “paleolithic” genes + modern diets

Nutrigenomics Quantification of the nutritional genotype-

phenotype

Phenotype

Metabolome

Proteome

Transcriptome

Epigenome

Genotype

Lifestyle

Nutrition

Environment

1 Genotype => 5 nutritional phenotypes

155 kg 76 kg

VLDL LPL

Remnant

FFA

Lipids

Chylomicrons

Organ and systemic responses to dietary lipids

Understanding NutritionHow nutrients regulate our genes: via sensing molecular

switches

Improved organ

capacity by PUFAs

J Clin Invest. 2004;114:94-103J Biol Chem. 2006;28:934-44 Endocrinology. 2006;147:1508-16Physiol Genomics. 2007;30:192-204Endocrinology. 2007;148:2753-63 BMC Genomics 2007; 8:267 Arterioscler Thromb Vasc Biol. 2007;27:2420-7

Am J Clin Nutr. 2007;86(5):1515-23PLOS ONE 2008;3(2):e1681 BMC Genomics 2008; 9:231BMC Genomics 2008; 9:262J Biol Chem. 2008;283:22620-7Arterioscler Thromb Vasc Biol. 2009;29:969-74.Plos One 2009;4(8):e6796HEPATOLOGY 2010;51:511-522

Am J Clin Nutr. 2009; 90:415-24Am J Clin Nutr. 2009;90:1656-64Mol Cell Biology 2009;29:6257-67Am J Clin Nutr. 2010;91:208-17BMC Genomics 2009Physiol. Genomics 2009Circulation 2010Diabetes 2010Cell Metabolism 2010Nature 2011

LPL

CE/TG

Chylomicron

CE/TG

Chylomicron remnant

FFA

Angptl4

Kersten, S. et al. Arterioscler Thromb Vasc Biol 2009;29:969-974

Expression profile of ANGPTL4 mRNA in human tissues

Angptl4-\- mice on HFD become very ill

Lichtenstein et al. Cell Metabolism 2010

Inflammatory response independent of microbiota

Lichtenstein et al. Cell Metabolism 2010

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

Lichtenstein et al. Cell Metabolism 2010

No effect of medium chain or PUFA TGs

Lichtenstein et al. Cell Metabolism 2010

Angptl4 protects against lipolysis and subsequent foam cell formation

Angptl4 protects against lipolysis and subsequent foam cell formation

Adipocytes at the crossroads of energy homeostasis

Normal

Type 2 Diabetes

Visceral Fat Distribution:Normal vs Type 2 Diabetes

Metabolic defects leading to the development of hepatic steatosis

Metabolism & Inflammation

Liver, FAT & NASH/NAFLD

Nonalcoholic Fatty Liver Diseases (NAFLD): Liver component of Metabolic Syndrome

Different stages in NAFLD progression:

Molecular events involved in NASH pathogenesis: Role of PPARa (Endocrinology 2008 & Hepatology 2010) Role Kupffer cells (Hepatology 2010)

Role of macrophages in lipid metabolism (JBC 2008; Cell Metabolism 2010)

hepatic steatosis steatohepatitis (NASH) & fibrosis cirrhosis

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.

Experimental Design

• stratificatio

n on body weight

• liver• plasma collection

multiple protein assays

RNA extraction: Affx microarrays

tissue collectionrun-in diet 20 weeks diet intervention

frozen sections: histological feat.

• ep. white adipose tissue

10% low fat diet

(palm oil)

10 LFD

10 HFD

45% high fat diet

(palm oil)

20 LFD

RNA extraction: real-time PCR

paraffin sections: histological feat.

lipid content

quality control & data analysis

pipeline

Mouse genome 430 2.0

0 2 4 8 12 16 20 weeks-3

High fat diet-induced obesity

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weeks under diet intervention

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LFLLFH

HFLHFH

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Liver TG content

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LFL LFH HFL HFH

Adipose dysfunction in HFH mice

Leptin

A subpopulation of mice fed HFD develops NASH

Immunohistochemical staining confirms enhanced inflammation and early fibrosis in HFH mice

Macrophage CD68

Collagen

Stellate cell GFAP

Results I

• Mice exhibited pronounced heterogeneity in liver histological scoring, leading to classification into 4 subgroups: – LF-low (LFL) responders displaying normal liver morphology, – LF-high (LFH) responders showing benign hepatic steatosis, – HF-low (HFL) responders displaying pre-NASH with

macrovesicular lipid droplets, – HF-high (HFH) responders exhibiting overt NASH characterized

by ballooning of hepatocytes, presence of Mallory bodies, and activated inflammatory cells.

Upregulation of inflammatory and fibrotic gene expression in HFH

responder mice

Adipose dysfunction in HFH mice

Change in adipose gene expression indicate adipose tissue dysfunction

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

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.

Diabetes. 2010;59:3181-91.

Human applications?Individual protein profiles

Population I (MARIS, n=56)

Van Dijk et al. Plos One 2010

Design of the SFA vs MUFA-rich intervention study

Run-in

SFA-rich diet (n=20)

SFA-rich diet (n=10)

MUFA-rich diet (n=10)

Baseline- Clamp- Adipose tissue

biopsy- Blood sampling

After intervention- Clamp- Adipose tissue

biopsy- Blood sampling

T=0 wks T=2 wks T=10 wks

Van Dijk et al. AJCN 2009

‘Obese-linked’ pro-inflammatory gene expression profile by SFAs

• 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”

SFA diet MUFA diet

Van Dijk et al. AJCN 2009

Fish-oil supplementation induces anti-inflammatory gene expression profiles in human blood

mononuclear cells

Less inflammation & decreased pro-arteriosclerosis markers= Anti-immuno-senescence

Bouwens et al. Am J Clin Nutr. 2009

Summary

• Less healthy: Dietary fats rich in long chain saturated fatty acids that can be pro-inflammatory if chronically “overconsumed”

• More favourable: Unsaturated fatty acids (in particular PUFAs from fish oil) have anti-inflammatory properties

• A healthy adipose tissue is essential to efficiently store fat and prevent ectopic fat deposition

• Healthy : Subcutanous fat > visceral fat > ectopic fat : Unhealthy

• Future challenge: To prevent the unhealthy effects of a surplus of added sugars (sucrose, fructose) & high GI carbs– Will be converted into saturated fat– Linked to ectopic fat deposition e.g. NASH– Linked to obesity, diabetes, CVD….– Childhood obesity

Thanks

Lydia AfmanMark BouwensSusan van DijkDiederik EsserSergio Lopez

Lisette de GrootMarianne GeleijnseOndine van de RestMarieke BosEdith FeskensRik HeijligenbergDianne HoelenJeanne de VriesGeert Heidema