Charles C. Muscoplat, PhDVice President for Statewide

StrategicResource Development

McKnight Presidential Endowed Chair

Professor of Medicine and Food Science and Nutrition

NutrigenomiNutrigenomicscs::

Genes in Your Food Genes in Your Food and Genes in Youand Genes in You

Sponsored Projects SymposiumFebruary 13, 2008

Goals TodayGoals Today• Be able to understand how foods and/or nutrients can

affect our health by mechanisms other than as providing simple nutrients, vitamins or energy

• Illustrate that some foods or food products can treat and/or prevent major human diseases other than by providing “adequate” or essential nutrition or energy by altering gene expression or through existing genetic polymorphisms

• Demonstrate that we can bring together agriculture as a full partner in human health promotion and disease prevention by utilizing foods as Medicine “ala Hippocrates”.

• To better establish that we can use foods to alter or complement gene expression in a dose and time dependent manner to prevent, ameliorate or cure serious human diseases.

Diet and Chronic Diet and Chronic DiseasesDiseases

• 125 million Americans have 1 or more chronic disease conditions (e.g., coronary heart disease, diabetes, cancer, stroke, mental illness) that may be related to diet and/or lifestyle factors

• Chronic diseases account for 75% of all health care expenditures

• Costs for chronic diseases is approaching $1 trillion

• Modifiable factors; diet, lifestyle, exercise, alcohol, tobacco

• Not modifiable factors; age, sex, geneotype, family history

• Many of these diseases and conditions can be prevented or treated with modifiable changes in diet and lifestyle: – GENES IN YOUR FOOD - GENES IN YOU

Today’s Medicine and Nutrition. Today’s Medicine and Nutrition. One size does not fit all. Not One size does not fit all. Not

everyone responds similarly to everyone responds similarly to medications or food:medications or food:

Patients are “Different” by Patients are “Different” by PolymorphismsPolymorphisms

Oranges, Grapefruits, Mandarin Orange, Lemons, Limes, Citrons, Shaddocks, Pummelos, Osbecks, Sour Oranges. Do they share common genes or are some genes on or off? Which alleles are healthful and which are not?

Genetic Variability Exists in our Food: Alleles, Regulation, EpigeneticsIf you want health, which varieties do you choose? Why?

Tomatoes, Cherry tomatoes, yellows, heirloom, fried green, Roma, Sun Gold, Big Boy, Plum, Purple. Is yellow lycopene as healthy as red lycopene?

Broccoli and Cauliflower share ancestral genes of origin, including nutrient constituent genes, but are they equally healthful to you? Which on is more healthful?

Soybeans have several important nutrients but levels vary among varieties. Ancient varieties have higher levels of cancer preventing compounds than modern varieties.

Choose a Tomato for your own purpose

Are these different?

Do they have different genes?

Are different genes expressed?

Are they equally healthy?

Do they grow the same?

Do rabbits eat them?

To they taste the same?

Carrots?

Monarch caterpillars and butterflies are genetically identical and have

their genes “On or Off” Depending ?The concept of genomics is “Gene Expression”

Genetically identical - but differing in gene expression

Genomics and Body Plans:

Which of these plans is not like the others?

Which of these plans are kind of the same?

Learning about structural genomics and evolution

Learning about worms and flies can teach us about ourselves and our genes

Flies, worms and People have the same basic body plan origin – We all have 14 homeotic body plan gene segments:

3 head, 3 chest and 8 abdomen

Embryonic Development IllustratesConservation of genes over 600 million years

We all have the same basic body plan…look around you...

Homeotic Hox Genes describe all basic body plans

What is so unusual about this fly eye?It is growing on a fruit fly leg!

Fruit Fly Compound Eye

Fly eye stimulated to grow on fly leg by injecting mouse Homeotic “eyeless” genes into fly leg

Homeotic Genes Work Across Species and Time

NutrigenomicsThe science of nutrigenomics seeks to provide a molecular understanding for how common dietary chemicals (i.e., nutrients) affect health by altering the expression and/or conformational structures of an individual’s genetic makeup.

Example 1: These two genetically identical mice were born of genetically identical

mothers who were fed differently in pregnancy and they will have very different

lives

Their identical mothers were fed different amounts of methylating nutrients or soy genistein during pregnancy

Yellow Mouse

Agouti Mouse

Cooney J Nutr 2002;132:2393S-2400S.

High risk cancer, diabetes, obesity & reduced lifespan

Lower risk of cancer, diabetes, obesity and prolonged life

Palindrome LTR HypomethylatedTransposon sequence

Palindrome LTR HypermethylatedTransposon sequence

Maternal Supplements

WithGenistein

zincmethionine

betainecholine,folate

B12

Epigenetics Occurs

Increasing MethylationChange in coat color

Change to lower lifetime weightChange to improved lifetime health

Increasing soy supplement genistein alters gene expression and thus phenotype

PNAS November 14, 2006 Vol. 103 no. 46 17-71-17072PNAS November 14, 2006 Vol. 103 no. 46 17-71-17072

What you eat or what your mothers ateWhat you eat or what your mothers ate

The role of the environment has been underplayed in developmental biology – Developmental Plasticity and Epigenetics

Developmental biology largely ignores “nongenetic” causes of individual variation. Yet, it is clear Yet, it is clear that the environment can exert a strong influence on development; e.g, developmental plasticity. that the environment can exert a strong influence on development; e.g, developmental plasticity. Although most evolutionary biologists recognize the environment as an important source of individual variation, many regard environmental responsiveness as developmental “noise” that has no long-term evolutionary consequences.

Clones of Daphnia

Reared in Reared in absence of absence of predatorspredators

Reared in presence Reared in presence of predators & of predators & predator kairomones predator kairomones DaphniaDaphnia develop develop spines, which deter spines, which deter predationpredation

Nutrient (Food) Methylation: Modulation of Gene- Environment and Gene-Diet

Interactions may be through DNA methylation

Methylation can alter genesMethylation can alter genesMethylation can alter genesMethylation can alter genesMethylation can alter genesMethylation can alter genesMethylation can alter genes

Nature’s way of allowing environmental factors to tweak gene expression without making permanent mutations. Primary DNA does not vary but they can be altered to read differently

Waterland and Jirtle at Duke University:

What your mother ate can determine your

lifetime outcome through epigenetic mechanisms?

…….. the metastable methylation status of specific transposable element insertion sites renders them epigenetically labile to early methyl donor nutrition. Our results show that dietary methyl supplementation of a/a dams with extra folic acid, vitamin B(12), choline, and betaine alter the phenotype of their A(vy)/a offspring via increased CpG methylation at the A(vy) locus and that the epigenetic metastability which confers this lability is due to the A(vy) transposable element. These findings suggest that dietary supplementation, long presumed to be purely beneficial, may have unintended deleterious influences on the establishment of epigenetic gene regulation in humans……………….

………….. Here, we report that maternal dietary genistein supplementation of mice during gestation, at levels comparable with humans consuming high-soy diets, shifted the coat color of heterozygous viable yellow agouti (A(vy/a) offspring toward pseudoagouti. This marked phenotypic change was significantly associated with increased methylation of six cytosine-guanine sites in a retrotransposon upstream of the transcription start site of the Agouti gene. The extent of this DNA methylation was similar in endodermal, mesodermal, and ectodermal tissues, indicating that genistein acts during early embryonic development. Moreover, this genistein-induced hypermethylation persisted into adulthood, decreasing ectopic Agouti expression and protecting offspring from obesity. Thus, we provide the first evidence that in utero dietary genistein affects gene expression and alters susceptibility to obesity in adulthood by permanently altering the epigenome……….

Dolinoy, Weidman, Waterland and Jirtle at Duke University

Pseudo-agouti predicts body weight throughout adulthood.

Geinstein (soy) supplementation increases the incidence of normal-body-weight animals as adults.

Timeline of methylation consequences.

Failure to methylate early results in the obese and sick mouse syndrome.

Very early dietary events can have lifelong consequences!

Eating 10 g to 40 g Soy NutsWhat if you need soybeans to treat your disease?If food can also be a biopharmaceutic, then eating could be viewed as ‘dosing’. e.g, Eat soy genistein to a steady dose of 2,000 nmol/Lh AUC. Eat to Cmax of 2,000 nmole/L; Eat to Signal Transduction then stop? There is a goal to eat healthy!

pK data T ½ for genistein ~ 10 hrs and AUC

Soy genistein can markedly augment cytotoxicity of certain anti-neoplastic agents

Soy genistein plus Cisplatin is profoundly antineoplastic; p< 0.01

25 uM Genistein as a bioharmaceutic agent; was the dose optimal; was the agent from soy optimal, was the timing optimal, was the purity high quality?

Soybean genistein inhibits NF kappa Beta in a dose dependent manner promoting apoptosis:

Cisplatin activates NF-kB to prevent apoptosis.

Thus the two agents are synergistic as anti-neoplastic agents

Soy genistein + Cisplatin caused tumor reduction and growth delay compared to either cisplatin alone or genistein alone. The effects are biologically complementary.

Soy isoflavones as biopharmaceutics. Should all men over age 50 consume soybean extracts of genistein at a dose determined to inhibit NF-Kappa beta and signal transduction in steady state? How about men with early lesions in individuals in ‘watchful waiting”. Risk benefit?

Soy isoflavones as cancer therapy

Log Level Rise of PSA Levels in Patients Taking Soy Supplements

But? Was the dosing optimal? This study used one capsule daily. The pk data would support at least bid dosing? We do know volume of distribution and penetration into prostate acidic secretions? There was no dose titration to side effects or optimal efficacy.

Should patients receiving radiation be pre-medicated with soy based genistein? What is risk benefit?

Alcohol Dehydrogenase has 3 alleles: only the ADH3 allele is associated with significantly reduced myocardial infarction in relationship to alcohol consumption. Women raised HDL to about 72 mg/dl while men raised HDL to about 51 mg/dl. HDL was increased in slow and intermediate alleles but MI data was not sig in these 2 groups.

Serum Triglycerides are influenced by both diet and genes

Change in diastolic blood pressure in response to fiber based diets with alleles

of AGT gene codon 235

-6

-5

-4

-3

-2

-1

0

1

2

T/T T/M M/M

Insoluble

Soluble

D

BP

(m

m H

g)

Hegele, et al. Nutr. Res. 17: 1229, 1997

TT Tt

Tt

tt

< 300 mg Caffeine > 300 mg

6

-12

0

-6

Vitamin D Receptor GenotypeTT Tt tt TT Tt tt

D S

pin

e B

on

e M

ass

Den

sity

(%

)Influence of caffeine on bone mass may depend on VDR allele

Rapuri et al. Am J Clin Nutr 2001 Nov;74(5):694-700

TTtt

Malformations caused by high doses of vitamin A, which was given to mothers on day 8 of pregnancy. Vitamin A has caused the homeotic HOX genes 1-4 to become expressed in groups of cells that usually do not express these genes.

Control of Homeotic Hox Gene Expression and Birth Defects: Vitamin A

Effect

Blood Pressure and Diet

F/V or Dash Diets can

Lower blood pressure

Only in individuals with AA genotype

but not GG Genotype.

Control of angiotensinogenRelated SBP and

DBPVia diet is gene-

related.

Jose M. Ordovas

CONCLUSIONS: We found a significant gene-diet interaction associated with the APOA1 G-A polymorphism. In women carriers of the A allele, higher PUFA intakes were associated with higher HDL-cholesterol concentrations, whereas the opposite effect was observed in G/G women = 8% PUFA

= 4-8% PUFA

= <4% PUFA

HD

L C

hole

ster

olAPOA1 G-A Polymorphism and PUFA Intake vs HDL

Pima Indians living on the Gila River Indian Reservation near Tucson Arizona have the highest rate of diabetes in the world, while Pimas in Maycoba, in the Mexican state of Sonora, rarely get the disease. Leslie Schultz has concluded that the differences in diet and and exercise may explain the contrast.

Pima Indians living on the Gila River Indian Reservation

-40

-30

-20

-10

0

10

20

ALL apo E2 apoE3 apoE4

TGLDL-CHDL-C

% change on fish oil

Minihane et al, 2000

Gene specific response to fish oil supplementation

The data demonstrate the efficacy of fish oil fatty acids in counteracting the proatherogenic lipid profile of the ALP but also that the apoE genotype influences responsiveness to this dietary treatment.

CONCLUSION: Intake of coffee was associated with an increased risk of nonfatal MI only among individuals with slow caffeine metabolism, suggesting that caffeine plays a role in this association.CONTEXT: The association between coffee intake and risk of myocardial infarction (MI) remains controversial. Coffee is a major source of caffeine, which is metabolized by the polymorphic cytochrome P450 1A2 (CYP1A2) enzyme. Individuals who are homozygous for the CYP1A2*1A allele are "rapid" caffeine metabolizers, whereas carriers of the variant CYP1A2*1F are "slow" caffeine metabolizers

Coffee or associated lifestyle?

Are coffee abstainers at risk?

Does risk depend on age?

Which component of coffee?

Is Coffee associated with CVD?

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

slow

rapid

CYP1A2

Genotype

Controls

%

Cases

%

A/A

A/C

C/C

46

43

10

45

44

11

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

Coffee Intake and Risk of MI

Total Population

Od

ds

Rat

io

0

1

2

3

4

<1 cup/d1 cup/d2-3 cups/d4+ cups/d

*

* P<0.05

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

CYP1A2 Genotype

*1A/*1A *1A/*1F + *1F/*1F

Od

ds

Rat

io

0

1

2

3

4

<1 cup/d1 cup/d2-3 cups/d4+ cups/d

A/A A/C + C/C

Coffee Intake and Risk of MI

* P<0.05

**

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

CYP1A2 Genotype

*1A/*1A *1A/*1F + *1F/*1F

Od

ds

Rat

io

0

1

2

3

4

<1 cup/d1 cup/d2-3 cups/d4+ cups/d

A/A A/C + C/C

Subjects <59 Years of Age

Coffee Intake and Risk of MI

* P<0.05

*

*

*

*

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

CYP1A2 Genotype

*1A/*1A *1A/*1F + *1F/*1F

Od

ds

Rat

io

0

1

2

3

4

<1 cup/d1 cup/d2-3 cups/d4+ cups/d

A/A A/C + C/C

Subjects <50 Years of Age

Coffee Intake and Risk of MI

* P<0.05

*

*

*

*

Coffee or associated lifestyle?Are coffee abstainers at risk?Which component of coffee?

Slide permission & courtesy of Ahmed al-SohemySlide permission & courtesy of Ahmed al-Sohemy

Genistein (soy) & Mammary Cancer:Timing is Everything

LaMartiniere et al JNutr 132: 552S, 2002

Exposure Period Tumors/Rat

None

Prenatal

Adult

PrepubertalPrepubertal

Prepubertal + AdultPrepubertal + Adult

8.9

8.8

8.2

4.34.3

2.82.8

Slide courtesy of Dr. J. Milner, NCI

If you don’t like it, you won’t eat it

What you like to eat is genetic: are you a genetic supertaster, genetic non-taster or moderate taster?

Goals TodayGoals Today• Be able to understand how foods and/or nutrients can

affect our health by mechanisms other than as providing simple nutrients or energy

• Illustrate that some foods or food products can treat and/or prevent major human diseases other than by providing “adequate” or essential nutrition or energy by altering gene expression or through existing genetic polymorphisms

• Demonstrate that we can bring together agriculture as a full partner in human health promotion and disease prevention by utilizing foods as Medicine “ala Hippocrates”.

• To better establish that we can use foods to alter or complement gene expressionin a dose and time dependent manner to prevent, ameliorate or cure serious human diseases.

The Fog of Nutrition: What can you believe? The public is at a complete loss to understand what to eat to stay healthy. The minute, hourly, daily, weekly, monthly contradictions on nutrition recommendations and “diets” recommendations are nearly irresponsible and misleading. Now is the time to provide careful, albeit complex recommendations based upon large prospective controlled clinical trials utilizing well established food biopharmaceteutics and individualized nutrition guidelines along with konwledge of genetic polymorphisms of foods and people

OF NUTRITION

Thank you…...