Nutrigenomics and Cardiovascular Disease

Brian J Bennett, PhDResearch Leader

Obesity and Metabolism UnitWestern Human Nutrition Research Center

USDA-Agricultural Research ServiceDavis,CA

Outline

• Introduction

• Candidate genes and nutri-genetic studies

• GWAS

• GWAS candidates and nutri-genetic

• Mouse studies

Why CVD?

• CVD is the leading cause of death in the US

• In 2010, total costs of CVD were estimated to be $444 billion.

• Treatment accounts for about $1 of every $6 spent on health care in the US.

National Epidemic of Metabolic Disease

1. As geneticist we know there are genetic factors

2. Clearly there are environmental factors

3. As nutrition scientist: diet is an important environmental factor

• To diagnose metabolic syndrome, most doctors look for the presence of three or more of these components:

• Central or abdominal obesity (measured by waist circumference):– Men - greater than 40 inches– Women - greater than 35 inches

• Triglycerides greater than or equal to 150 milligrams per deciliter of blood (mg/dL)

• HDL cholesterol:– Men - Less than 40 mg/dL– Women - Less than 50 mg/dL

• Blood pressure greater than or equal to 130/85 millimeters of mercury (mmHg)

• Fasting glucose greater than or equal to 100 mg/dL•

Diagnosis of Metabolic Disease

Differences between racial groups in metabolic syndrome

https://www.cdc.gov/pcd/issues/2017/16_0287.htm

• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2675814/

Metabolic syndrome-

complex interactions lead to diabetes and cardiovascular disease

Candidate genes in lipoprotein metabolism and atherosclerosis

Aldons J. Lusis et al. Circulation. 2004;110:1868-1873

Copyright © American Heart Association, Inc. All rights reserved.

Aldons J. Lusis et al. Circulation. 2004;110:1868-1873 Copyright © American Heart Association, Inc. All rights reserved.

Atherosclerosis is a complex disease involving multiple genes, cell types and metabolic

process at the vessel wall

Current Diet recommendations

Heart Healthy

As you make daily food choices, base your eating pattern on these recommendations:

•Eat a variety of fresh, frozen and canned vegetables and fruits without high-calorie sauces or added salt

and sugars. Replace high-calorie foods with fruits and vegetables.

•Choose fiber-rich whole grains for most grain servings.

•Choose poultry and fish without skin and prepare them in healthy ways without added saturated

and trans fat.

•Eat a variety of fish at least twice a week, especially fish containing omega-3 fatty acids (for example,

salmon, trout and herring).

•Select fat-free (skim) and low-fat (1%) dairy products.

•Avoid foods containing partially hydrogenated vegetable oils to reduce trans fat in your diet.

•Limit saturated fat and trans fat and replace them with the better fats, monounsaturated and

polyunsaturated.

•Cut back on beverages and foods with added sugars.

•Choose foods with less sodium and prepare foods with little or no salt

•If you drink alcohol, drink in moderation. That means no more than one drink per day if you’re a woman

and no more than two drinks per day if you’re a man.

•Follow the American Heart Association recommendations when you eat out, and keep an eye on

your portion sizes.

http://www.heart.org/HEARTORG/HealthyLiving/HealthyEating/Nutrition/The-American-Heart-Associations-Diet-and-Lifestyle-Recommendations_UCM_305855_Article.jsp#.WSJYyWjys2w

Why PUFA’s?

Potential cardio-protective mechanisms of PUFA’s

PUFA’s are generally anti-inflammatory

Potential Gene x Diet interactions

Generalized pathway for the conversion of eicosapentaenoic acid to eicosanoids.

Philip C Calder Am J Clin Nutr 2006;83:S1505-1519S

PUFA’s are generally anti-inflammatory

Genotype guided intervention

PCR

DNA isolation

Participant recruiting

Randomized Control Trial for NGx interactions

Charles B. Stephensen et al. J. Lipid Res. 2011;52:991-1003

©2011 by American Society for Biochemistry and Molecular Biology

20 AA females

dd dd5d 55

20 AA females

20 AA females

5d 55

20 AA females

20 AA females

20 AA females

Placebo60 AA females

1 gm Fish oil concentrate 60 AA females

five Sp1 element tandem re- peats]

ALOX5PromoterVariants

Mean (±SEM) changes in oxylipid concentrations are shown from baseline to follow-up among the

three ALOX5 genotype groups of interest in response to placebo and fish oil interventions.

Charles B. Stephensen et al. J. Lipid Res. 2011;52:991-

1003

Randomized Control Trial for NGx interactions

Summary of findings

• this placebo-controlled intervention trial, randomized within genotypes,

• identified a significant gene–diet interaction• genotype-determined differences in

response to omega-3 intake

Outline

• Introduction

• Candidate genes and nutri-genetic studies

• GWAS

• GWAS candidates and nutri-genetic

• Mouse studies

Published GWA Reports, 2005 – 6/2012To

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Calendar QuarterThrough 6/30/12 postings

0

200

400

600

800

1000

1200

1400

2005 2006 2007 2008 2009 2010 2011 2012

1350

Published Genome-Wide Associations through 07/2012Published GWA at p≤5X10-8 for 18 trait categories

NHGRI GWA Catalogwww.genome.gov/GWAStudieswww.ebi.ac.uk/fgpt/gwas/

Cardiogram results

Initial GWAS Finding

R McPherson et al. Science 2007;316:1488-1491Published by AAAS25

Initial GWAS Finding

GNET/NUTR 865

Fine mapping of the genomic interval on chromosome 9 associated with CHD. (A) SNPs spaced ∼5 kb apart in the

interval extending 175 kb upstream and downstream of rs10757274 and rs2383206 were assayed in 500 cases and

500 controls from the OHS population with GeneChip Human Mapping 500K Array Sets (Affymetrix, Santa Clara, CA).

R McPherson et al. Science 2007;316:1488-1491

9p21 Locus

9p21 locus- Associations reported

Disease/Trait Reported Gene(s)Strongest SNP-Risk Allele P-value # pubs

Abdominal aortic aneurysm CDKN2A,CKDN2B rs2383207-G 2 x 10-8 1

Ankle-brachial index CDKN2B rs10757269-G 9 x 10-9 1

Breast cancer CDKN2A,CDKN2B rs1011970-T 3 x 10-8 1

Coronary artery calcification CDKN2B rs1333049-C 3 x 10-24 1

Coronary heart disease CDKN2B-AS1 rs4977574-G 2 x 10-25 7

Endometriosis CDKN2BAS rs10965235-C 6 x 10-12 2

Glaucoma CDKN2B-AS1 rs2157719-? 2 x 10-18 5

Glioma CDKN2A, CDKN2B rs2157719-? 5 x 10-16 4

Intracranial aneurysm CDKN2A,CDKN2B rs1333040-T 2 x 10-22 4

Melanoma CDKN2A rs7023329-A 4 x 10-7 2

Myocardial infarction CDKN2A,CDKN2B rs4977574-G 3 x 10-44 2

Nasopharyngeal carcinoma CDNK2A,CDKN2B rs1412829-? 5 x 10-7 1

Platelet counts CDKN2A rs3731211-A 6 x 10-14 1

Type 2 diabetes CDKN2A, CDKN2B rs2383208-A 2 x 10-29 12

http://www.genome.gov/gwastudies/index.cfm

Kelly A. Frazer, Sarah S. Murray, Nicholas J. Schork & Eric J. TopolNature Reviews Genetics 10, 241-251 (April 2009)

9p21.3 locus- structure varies by population

Can 9p21 Risk Be Modified by Dietary Intake?

The Effect of Chromosome 9p21 Variants onCardiovascular Disease May Be Modified by DietaryIntake: Evidence from a Case/Control and a ProspectiveStudy

Ron Do1, Changchun Xie2,3, Xiaohe Zhang2, Satu Mannist o4, Kennet Harald4, Shofiqul Islam2,3,

Swneke D. Bailey1, Sumathy Rangarajan2, Mat thew J. McQueen2, Rafael Diaz5, Liu Lisheng6, Xingyu

Wang7, Kaisa Silander4,8, Leena Peltonen4,8{ , Salim Yusuf2, Veikko Salomaa4, James C. Engert1,9,10*,

Sonia S. Anand2,3*, on behalf of the INTERHEART invest igators

1 Department of Human Genetics, McGill University, Montreal, Quebec, Canada, 2 Population Health Research Institute, Hamilton Health Sciences, and McMaster

University, Hamilton, Ontario, Canada, 3 Departments of Medicine and Clinical Epidemiology and Biostatistics, McMaster University, Hamilton, Ontario, Canada,

4 Department of Chronic Disease Prevention, National Institute for Health and Welfare, Helsinki, Finland, 5 Estudios Cardiologicos Latinoamerica, Rosario, Argentina,

6 Cardiovascular Institute and Fu Wai Hospital, Chinese Hypertension League Institute, Beijing, China, 7 Laboratory of Human Genetics, Beijing Hypertension League

Institute, Beijing, China, 8 Human Genomics Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Helsinki, Finland, 9 Department of Medicine,

McGill University, Montreal, Quebec, Canada, 10 The Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada

Abst ract

Background: One of the most robust genetic associations for cardiovascular disease (CVD) is the Chromosome 9p21 region.However, the interaction of this locus with environmental factors has not been extensively explored. We investigated theassociation of 9p21 with myocardial infarction (MI) in individuals of different ethnicities, and tested for an interaction withenvironmental factors.

Methods and Findings: We genotyped four 9p21 SNPs in 8,114 individuals from the global INTERHEART study. All fourvariants were associated with MI, with odds ratios (ORs) of 1.18 to 1.20 (1.856 102 8# p# 5.216 102 7). A significant interaction(p = 4.06 102 4) was observed between rs2383206 and a factor-analysis-derived ‘‘prudent’’ diet pattern score, for which amajor component was raw vegetables. An effect of 9p21 on MI was observed in the group with a low prudent diet score(OR= 1.32, p = 6.826 102 7), but the effect was diminished in a step-wise fashion in the medium (OR= 1.17, p = 4.96 102 3) andhigh prudent diet scoring groups (OR= 1.02, p = 0.68) (p = 0.014 for difference). We also analyzed data from 19,129individuals (including 1,014 incident cases of CVD) from the prospective FINRISK study, which used a closely related dietaryvariable. In this analysis, the 9p21 risk allele demonstrated a larger effect on CVD risk in the groups with diets low or averagefor fresh vegetables, fruits, and berries (hazard ratio [HR] = 1.22, p = 3.06 102 4, and HR= 1.35, p = 4.16 102 3, respectively)compared to the group with high consumption of these foods (HR= 0.96, p = 0.73) (p = 0.0011 for difference). Thecombination of the least prudent diet and two copies of the risk allele was associated with a 2-fold increase in risk for MI(OR= 1.98, p = 2.116 102 9) in the INTERHEART study and a 1.66-fold increase in risk for CVD in the FINRISK study (HR= 1.66,p = 0.0026).

Conclusions: The risk of MI and CVD conferred by Chromosome 9p21 SNPs appears to be modified by a prudent diet highin raw vegetables and fruits.

Please see later in the article for the Editors’ Summary.

Citat ion: Do R, Xie C, Zhang X, Mannisto S, Harald K, et al. (2011) The Effect of Chromosome 9p21 Variants on Cardiovascular Disease May Be Modified by DietaryIntake: Evidence from a Case/Control and a Prospective Study. PLoS Med 8(10): e1001106. doi:10.1371/journal.pmed.1001106

Academic Editor: Cathryn Lewis, Kings College London, United Kingdom

Received January 11, 2011; Accepted August 30, 2011; Published October 11, 2011

Copyright : ß 2011 Do et al. This is an open-access article distributed under the terms of the Creative Commons Attribut ion License, which permits unrestricteduse, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This study was funded by the Heart and Stroke Foundation of Ontario (Grant # NA6336). SA holds the Heart and Stroke Foundation of Ontario MichaelG. DeGroote endowed Chair in Population Health, and the May Cohen Eli Lilly endowed Chair in Women’sHealth at McMaster University. VSwas supported by theFinnish Foundation for Cardiovascular Research, Sigrid Juselius Foundation, and the Academy of Finland (grant number 129494). SY is supported by an endowedchair from the Heart and Stroke Foundation of Ontario. RDo is a recipient of a Frederick Banting and Charles Best Canada Graduate Scholarship Doctoral Awardfrom the Canadian Institutes for Health Research. SDB holds a graduate scholarship from the McGill University Health Centre Research Institute and theDepartment of Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compet ing Interests: The authors have declared that no competing interests exist.

Abbreviat ions: CVD, cardiovascular disease; FFQ, food frequency questionnaire; HR, hazard ratio; LD, linkage disequilibrium; MI, myocardial infarction; OR, oddsratio

* E-mail: jamie.engert@mcgill.ca (JCE); anands@mcmaster.ca (SSA)

{ Deceased.

PLoS Medicine | www.plosmedicine.org 1 October 2011 | Volume 8 | Issue 10 | e1001106

- Five ethnicities- Retrospective Case-Control study Risk factors for acute non-fatal MI- Recruitment within24 hours of being admitted to a coronary care unit with clinicalcharacteristics of acute MI. Control matched for age and sex.

Four SNPs (rs10757274, rs2383206, rs10757278, rs1333049) from the Chromosome 9p21region were selected based on previous results from genome-wide association studies forcoronary heart disease/MI

Genotypes : 1,744 Europeans, 1,867 South Asians, 2,231 Chinese, 1,100 Latin Americans, and 1,172 Arabs (a total of 8,114 samples).

Can 9p21 Risk Be Modified by Dietary Intake?

INTERHEART

The Effect of a Prudent Diet is most pronounced in

individuals with Risk Allele

An analysis of the rs2383206 genotype and tertiles of the prudent diet score in theINTERHEART samples demonstrated that individuals with two copies of the risk allele (GG)and with a low prudent diet score had a 2-fold increase in MI risk when compared to thereference group of individuals with two copies of the protective allele (AA) and a highprudent diet score.

Will Dietary interventions be most effective in these people

rs2383206

Conclusions

• Metabolic syndrome is complex and multigenic– Candidate genes regulate metabolism

– Diet is also important

• Genotype guided interventions– Demonstrate differences in metabolism

– Changes effect of diet

• Main effect alleles of GWAS demonstrate potential benefit effects of diet