More Sachin C.

NUTRIGENOMICS

"If we could give every individual the right amount of nourishment and exercise, not too little and not too much, we would have found the safest way to health.”

"Leave your drugs in the chemist's pot if you can heal the patient with food." - Hippocrates

Balanced Diet Pyramid

Family Health at Risk

Nutrients acts as dietary signals

Energy homeostasis

Energy homeostasis

Nutrient absorptionNutrient

absorption

Cell proliferation

Cell proliferation

Nutritional factors

Gene transcription

Transcription factors

What are dietary signatures/profiles? Patterns of gene expression, protein expression and metabolite production in response to particular nutrients or nutritional regimes can be viewed as 'dietary signatures

How can these dietary signatures been used in Nutrigenomics research? Nutrigenomics seeks to examine these dietary signatures in specific cells, tissues and organisms, and to understand how nutrition influences homeostasis. Dietary signatures can be used to identify (early) molecular biomarkers

Nutritional Genomics

• Nutrigenomics studies modulating effects of nutrients on gene structure and expression; related science of Nutrigenetics researches patients’ unique responses-caused by specific genetic variations-to macro- and micronutrients

(Desiere, 2004)

• “… the interface between the nutritional environment

and cellular/ genetic processes” (Kaput and Rodriguez, 2004)

• Next frontier in the post genomic age

• Off shoot of HGP

Health effects of the nutrient and non-nutrient components of food relates specific molecular interactions

(Ommen et al., 2004)

Two sub definitions

• Two sides of a coin

• Dynamic cause–effect relationship between nutrition and the human genome

(Mutch et al., 2005)(Mutch et al., 2005)

Nutrigenomics Nutrigenetics

Aims to study the genome- wide influence of nutrition

Concerned with the functional effect of nutrient or non-nutrient food components on the genome, transcriptome, proteome and

metabolome

(Muller and Kersten, 2003)

Concerned with how genetic variation determines an individual’s disease risk, nutrient requirements, metabolic response and responsiveness to a bioactive dietary components or nutritional therapy

(DeBusk et al., 2005)

Two-way interaction between nutrition and the human genome

(Roche, 2004)(Roche, 2004)

The direct and indirect effects of nutrients

(Muller and Kersten, 2003)(Muller and Kersten, 2003)

Challenge

• Common, polygenic, diet-related diseases which are composite of multiple genetic variants interacting with multiple environmental factors

• To develop and apply a robust nutritional genomics research approach that is sensitive enough to take account of both genetic heterogeneity and diverse nutrient exposure

Nutrigenomics – A Systems Biology Approach

Bioactive food components influence genetic and epigenetic events

Dietary factors and the regulation of DNA methylation

“omics” of nutrition to identify how dietary factors contribute to phenotype

Implications of approach

• New way of dealing with nutrigenomics data aims to integrate all of the gene expression and/or proteomic data and present this information as a ‘complete’ biological process

• Multivariate analytical approach

• Key to our understanding of nutrition and health

Health Effect of Dietary FattyAcids and TFs

• Fatty acids had important metabolic effects with respect to energy homeostasis, lipoprotein metabolism, glucose homeostasis and inflammation

• Potent cellular molecules that interact with several transcription factors, which in turn explains the diverse health effect of altered dietary fat intake

Lipid sensitive transcription factors

Cross talk between lipid sensitive TFs

inhibits

LXR binding

regulation

Expression of SREBP-1c & other downstream lipogenic genes

regulationregulationregulationregulationregulation

PPARα activation

regulationregulationregulationregulation

(Clarke et al., 1999)(Clarke et al., 1999)

Metabolomics – The Newest Tool

• Utilizes analytical chemistry technologies, 1) NMR

2) MS

• Help Capture Data On Complete Metabolome

• May provide the comprehensive biomarker of multiple metabolites to asses nutrient status metabolic responses and disease predispositions

Metabolism approach

• n-6 PUFA and n-3 PUFA interventions

• Hepatic transcriptome and lipid metabolome completed

• Diets supplemented with AA or EPA or fish oil

• Studied down-regulation of hepatic SCD1 and not SCD2 or SCD3

• SCD1 potential therapeutic target to protect against CVD and diet-induced obesity

Nutrigenetics – Individual GeneticVariability And Responsiveness

• Human genome 99.9% identical

• 0.1% variation principally due to SNPs, responsible for 90% variation among individuals

• Functional and Non-functional SNPs

• Gene-Nutrient interaction concept important

• Standard “one-size-fits-all” strategy is no longer considered to be good practice

Interrelationships Among Specific Nutrients,Genes and Health Outcomes

• Green Tea (Camellia sinensis)- Potent antioxidants-prevent certain cancers and

heart disease- Some women appear to show a reduction in

breast cancer- Have gene variant that produces less active form

of gene enzyme COMT inactivating carcinogenic compounds

• Curcumin: -yellow pigment from turmeric (Curcuma longa) - basic ingredient in Curries -produce suppressing genes for enzymes that produce inflammatory prostaglandins e.g. cyclooxygenase-2 (COX-2)• Prostaglandins linked to colon cancer, heart

disease, arthritis, and Alzheimer’s disease

• “curry-eating people of India have the world’s lowest incidence of Alzheimer’s disease”

- Sally A. Frautschy, Professor of Neurology,

University of California

Contd…

PERSONALIZED NUTRITION

Senior Scientist Jose OrdovasTufts' Human Nutrition

Research Center on Aging

• Pioneer worker in Nutrigenomics

• Talks about virtual lab

• Personalized nutrition for everyone in cost effective manner and behavioral guidelines for each individual depending on ones health status to reduce disease predisposition

• To be cautious regarding interpretations we need to move step-by-step like solving puzzle to create big picture

Step 1Step 1

Collecting (patient) informationCollecting (patient) information

• Physical examinationPhysical examination

• QuestionnaireQuestionnaireDietDietDisease historyDisease historyLifestyleLifestyle

Step 2Step 2

Looking for genetic differences that mirror physical Looking for genetic differences that mirror physical differences in patientsdifferences in patients

– PCRPCR– Gel electrophoresisGel electrophoresis– DNA sequencingDNA sequencing

Step 3Step 3

Examining the foodsExamining the foods

You can separate the different chemicals that make You can separate the different chemicals that make up a food using their different properties by:up a food using their different properties by:

– Different solubilityDifferent solubility– ChromatographyChromatography

Step 4Step 4

Test the effect of the food components on cellsTest the effect of the food components on cells

• Cell lines (luminescence)Cell lines (luminescence)

Step 5Step 5

Test the food components of the cells Test the food components of the cells in vivoin vivo

• Animal models (micro-arrays)Animal models (micro-arrays)

What happens next?What happens next?

• Make individualised dietary recommendations to Make individualised dietary recommendations to patients?patients?

• Produce dietary supplements to enhance individual Produce dietary supplements to enhance individual genetic potential?genetic potential?

• Produce dietary supplements to suppress gene Produce dietary supplements to suppress gene activity?activity?

Commercialization

• http://www.mycellf.com/MyCellfProgram.aspx

• Sciona Inc. (www.sciona.com)• Genelex Inc. (www.genelex.com)

CASE STUDIES

INTRODUCTION

• (PPARs), α and Ƴ ligand-activated transcriptional factors regulating lipid and lipoprotein metabolism; glucose homeostasis, and inflammation

• PPAR α- Expressed in liver -promotes microsomal ω-oxidation and peroxisomal β-

oxidation of Fatty acids Ligands- Fatty acids and FFs• PPAR Ƴ- Expressed in Adipose tissue -Ligands- UFAs• Isohumulones generated from humulones in Hops imparting bitter

taste to beer• Dual Agonists of both PPAR TFs (Yajimi et al.,

2004)

Research Methodology

• Materials: IHE by extracting hops with supercritical co2;

isomerised in alkaline condition • Animals and Diets:

-C57BL/6NCrj and PPAR α-/- /129S4/SvJae mice

-AIN-76A Diet• DNA Microarray analysis:

-RNA extraction

-cDNA labelling Cy3 or Cy5• QPCR: primers designed with primer3 software• Statistical Analysis: One way ANOVA

Results and Discussion

Biochemical parameters in male mice

Effect of IHE and FF :

Plasma Total cholesterol HDL cholesterol

Triglycerides Non esterified FFA

Transcriptional profiling and QPCR analyses in male C57BL/6N mice

• 8245 genes analysed out of which approx.1200 shown enough signal intensity

• IHE-20 and FF-100 genes: 2 fold or more expression

• No significant difference when diet supplemented

with Cholesterol

Dose-dependent induction of representative PPARα target genes in liver, by IHE

Gender differences in IHE-induced hepatic gene expression.

Conclusion

• Data showed IHE increased the expression of hepatic PPARα target genes

• Effect similar to FF• IHE modulated blood lipid status by

activating PPARα• So IHE may be potentially therapeutic for

preventing T2DM and Atherosclerosis• Beer consumption might actually help to

prevent alcoholic liver disease ‼

Introduction

• UVA exposure cause skin aging by singlet oxygen (1O2)-dependent pathways

• Gene regulation through TF AP-2.• β-carotene (βC) can protect skin since:

1. 1O2 quencher

2. scavenges ROS

3. Mildly reduces sunburns• HaCaT human keratinocytes used.(2 d, 1.5 µM)

Research Methodology

• βC and UVA Treatment of keratinocytes• Affymetrix GeneChip Analysis:

Five independent, factorially designed cell irradiation experiments analyzed by microarray hybridization.

• QRT-PCR to confirm key gene regulations in three independent cell irradiation experiments at different βC concentrations.(0.5, 1.5 and 3 µM)

• Apoptosis Assay : Caspase 3-activity 5 h after irradiation using CaspACE Assay System

Results and Discussion

• Cellular uptake of βC confirmed by HPLC• 1458 genes significantly regulated by at least one

treatment:

a) βC regulated 381 genes

b) UVA influenced 568 genes

c) UVA / βC regulated 1142 genes

d) not regulated by single treatment 610 genes

βC effects in unirradiated keratinocytes

Key process-skin aging

Marker genes

Receptor innate immunity

Differential regulation

Shown RA-independent effects

Genes up regulated

Genes down regulated

Down regulation

βC concentration dependently induced caspase-3 activity in

ultraviolet light A (UVA)-irradiated keratinocytes.

βC promoted keratinocyte differentiation but not

terminal steps

Ultraviolet light A (UVA) effects in keratinocytes

Differentially regulated proinflammatory genes

Enhanced tanning supplemented UVA

βC effects in UVA-irradiated keratinocytes

Quenched by βC

βC supplementation may indeed reduce wrinkling. (Battistutta et al., 2000)

Contd…

• RA target genes down regulated by UVA• Further promoted differentiation in irradiated

keratinocytes• This may render combined UVA / βC treatment a

promising therapy for skin disorders associated with disturbed differentiation. e.g. Psoriasis.

Proposed relationship of the modes of action of β -carotene to its influence on ultraviolet

light A-induced biological processes.

Conclusion

Discussion