Nutrigenomics: how food influences health how food influences health ... Epigenetics: The ability to ... Fish-oil supplementation for 6 months reduces

Nutrigenomics:

how food influences health

Dr. Katia Petroni

Dipartimento di Bioscienze

Università degli Studi di Milano

DIPARTIMENTO DI BIOSCIENZE

Large epidemiological studies

revealed that

the “Mediterranean diet”

significantly reduced

the risk of cardiovascular disease

(about -25%),

of the same order of magnitude as

pharmacological interventions

Higher consumption

of whole grain cereals

was also correlated with lower incidences

of not only cardiovascular disease,

but also obesity and type 2 diabetes

The link between diet and health is documented by

several epidemiological studies

Why

are fruits and vegetables

so important for our health?

Nutrients in plant foods

• Macronutrients • Proteins

• Oils

• Carbohydrates

• Fibre

• Micronutrients • Vitamins

• Minerals

• Phytonutrients • Alkaloids

• Polyphenols

• Carotenoids • Organosulphur compounds

• Nitrogen-containing compounds

Many phytonutrients possess biological activity

and are defined as bioactive compounds


Curcumin from turmeric Cyanidin 3-glucoside,

an anthocyanin from

blueberry and cherry

Quercetin, a flavonol

from apples and

onions

Genistein,

an isoflavone from

soy

Epigallocatechin

gallate (EGCG) from

green tea

Examples of bioactive compounds in plant foods

Polyphenols

Resveratrol

from red grape


Nutrigenomics studies the interaction of nutrients with the human

genome and aims to understand how nutrients influence gene

expression, metabolism and as a consequence human health

Nutrigenomics

Food Health

Genomics

Nutrigenomics

Nutrigenomics is the application of “omic” technologies to nutrition


Mechanisms

Pathways

Target Genes

Signatures

Profiles

Biomarkers

Foods

Nutrition

Transcriptomics

Proteomics

Metabolomics

Identification of

• dietary signals

• nutrient sensors

• signaling pathways

• target genes

Research goals of nutrigenomics

Transcriptomics

Proteomics

Metabolomics

Identification of

• dietary signatures

• biomarkers

for early diagnosis


How does food influence health?

Mechanisms? Dietary signals Macronutrients

Vitamins and Minerals

Phytonutrients

Health Outcomes

Health

Longevity

Chronic disease

Epigenetics

Gene expression

Enzyme modifications

Antioxidants

Anti-inflammatory


Epigenetics: The ability to change gene activity —without changing gene

sequence (no mutations involved).

Typically, epigenetic modifications include

- DNA methylation (gene silencing) and

- Histone acetylation (gene activation)

What does “epigenetics” mean?

Genetically identical individuals may have different gene expression and phenotypes due to

epigenetic modifications imposed by environmental cues (e.g. lifestyle and dietary habits)

Non-smoker Smoker


DNA methylation

Methylating the cytosine of a CpG

motif results in gene silencing and

silent heterochromatin

Nutrients


Epigenetic events during development

Gluckman et al. Nat. Rev. Endocrinol. 5, 401–408 (2009) DIPARTIMENTO DI BIOSCIENZE

Mother’s diet influences

fur color and health of progeny

Agouti

Mother’s diet

during gestation

Standard

diet

Progeny

wild-type allele

Normal expression

Healthy

Brown color

Agouti IAP Standard

diet

Obesity

Tumours

Yellow color Unmethylated Avy allele

Agouti overexpression

DIPARTIMENTO DI BIOSCIENZE Dolinoy et al, Environ Health Persp 114:567-572, 2006

Agouti IAP

Methylated Avy allele

Normal expression

Healthy

Brown color Genistein-rich

diet

Dutch famine

Is DNA methylation affected by nutrition also in

human individuals during gestation?

DIGITARE NOME DIPARTIMENTO SECONDA RIGA NOME DIPARTIMENTO

The Dutch famine

5 months

May ‘45 October ‘44

2414 people aged 60 years examined

of which 912 interviewed and 741 with clinical examinations

Rations were 1000 to 500 Kcal per day for adults

Supplementary rations were given to childrens and pregnants

Painter et al, Repr Toxicol 20:345-352, 2005 DIPARTIMENTO DI BIOSCIENZE

P F1 F2 F1

Dutch famine

during early

gestation

Grandmothers Sons Daughters Grandchildren

Higher birth weight

Higher incidence of

Obesity

Type 2 diabetes

Atherosclerosis

Coronary heart disease

Breast Cancer

Alzheimer

Painter et al, Repr Toxicol 20:345-352, 2005

After 60 years, adults exposed to Dutch famine during early

gestation showed higher incidence of chronic diseases

Higher incidence of

Obesity

de Rooij, Roseboom BMJ Open 2013 Veenendal et al, Int J Obst Gynaecol 2013


DNA methylation levels were significantly different

for 6 out of 15 genes after famine exposure

Tobi et al, Human Mol Genet 18:4046-53, 2009

60 individuals


Lower methylation of IGF2

correlated with higher birth weight

Epigenetic programming in utero affects adult risk

factors and disease

Diet can alter the epigenetic state of the genome leading to dramatic

deprogramming or reprogramming of large numbers of genes in metabolic

and physiological pathways.

This may affect the incidence of adult age chronic diseases

such as type 2 diabetes, cardiovascular disease, neurodegenerative disease

and cancer.

Epigenetic modifications are reversible!


Link et al, Biochem Pharmacol 80:1771-1792, 2010

Foods contain many

phytonutrients with

epigenetic and chromatin

remodeling properties,

making nutritional

intervention a possible way to

“reprogram” the epigenome

to promote health and

prevent disease

Importance of diet on epigenetics


Oxidative

stress

Chronic

inflammation

Neurological

disorders

Cancers

Cardiovascular

disease

Metabolic disorders Bone, muscular, skeletal

diseases

Obesity

Chronic

inflammatory diseases

Alzheimer’s disease

Parkinson’s disease

Different types

of cancers

Atherosclerosis

Coronary disease

Cerebrovascular

disease

Heart failure

Cardiomiopathy

Fatty liver disease

Type 2 Diabetes

Kidney renal failure

Chronic obstructive

pulmonary disease

Psoriasis

Chronic pancreatitis

Inflammatory bowel

disease

Rheumatoid arthritis

Osteoporosis

Osteoarthritis

Cardiovascular disease

Type 2 Diabetes

Hypertension

Fatty liver disease

Cancer

Cumulative damage from oxidative stress and chronic

inflammation leads to chronic diseases

Oxidative stress occurs when production of RONS

exceeds cellular antioxidant defenses

Infections,

inflammation

UV-B, drugs

Unhealthy diet

OXIDATIVE DAMAGE

Lipids, proteins, DNA

Chronic diseases

and aging

Antioxidant

defenses

RONS

Healthy diet


Many dietary polyphenols are more effective antioxidants

in vitro than vitamins E or C

Rice-Evans et al. (1997), TIPS, 2, 152-159

Antioxidant Activity (TEAC mM)

Vitamins

Vitamin C 1.0 ±0.02

Vitamin E 1.0 ±0.03

Flavonols

kaempferol 1.3 ±0.08

quercetin 4.7 ±0.12

Anthocyanins

delphinidin 4.4 ±0.11

cyanidin 4.4 ±0.03

Hydroxycinnamic acids

chlorogenic acid 1.3 ±0.02

caffeic acid 1.3 ±0.01

ferulic acid 1.9 ±0.02

coumaric acid 2.2 ±0.06


However, low bioavailability and instability at physiological pH suggest

that anthocyanins may function as molecular signals

• Low bioavailability (<7%)

• Low stability at physiological pH

Anthocyanins

RONS

mainly

in GI tract

Direct antioxidant

activity

Endogenous

antioxidant defenses


2. About 30% reduction of infarcted

zone in ischemia-induced mice

Toufektsian et al., J Nutr 2008

3. Increased preischemic level of

reduced glutathione GSH

1. Feeding trial with anthocyanin-rich

blue corn on rats (12 mg/Kg/day)

r1 c1 R1 C1

Anthocyanin-rich diet protects heart from infarct by

increasing endogenous antioxidant defenses


Anthocyanins

RONS

Keap1

Nrf2

Keap1

Nrf2

ARE Antioxidant and

detoxifying genes

(gGCS, HO-1, … )

Cytoplasm Nucleus

ARE, Antioxidant Responsive Element

ANTIOXIDANT DEFENSE

UV radiation

Unhealthy diet

Infections

acute inflammation

How do anthocyanins promote

endogenous antioxidant defences?


Cassidy et al., (2013) Circulation 127(2):188-96

Perspective study on 93.600 women 25 to 42

years of age from the Nurses' Health Study II who

compiled food-frequency-questionnaires on their

dietary habits every 4 years for 18 years.

Women consuming >3 servings of

anthocyanin-rich foods (strawberries and

blueberries) per week showed a 32% lower

risk of myocardial infarction compared to

those consuming these fruits less than 1 time

per month.

Anthocyanin intake was 35 mg per day on average

(105 g strawberries or 21 g blueberries)


Acute inflammation is the immediate defensive reaction to infections or injuries

Vasodilation

Immune cell recruitment

RONS and interleukins

Resolution and health

in 2 weeks or less

If the inflammatory inducer

is not eliminated Immune cell infiltration

Increased production

of inflammatory mediators

Chronic inflammation

by the acute phase response

Acute and chronic inflammation

Medhhzitov, Cell 2010 DIPARTIMENTO DI BIOSCIENZE

Anthocyanins

Keap1

Nrf2

Keap1

Nrf2

ARE Antioxidant and

detoxifying genes

(gGCS, HO-1, … )

Cytoplasm Nucleus

ARE, Antioxidant Responsive Element

ANTIOXIDANT DEFENSE

NfkB

kB Pro-inflammatory

genes

(COX-2, IL-6, IL-8, … )

INFLAMMATORY RESPONSE

NfkB

IkB

kB, NfkB Binding Site

UV radiation

Unhealthy diet

Infections

inflammation

RONS

RONS

UV radiation

Unhealthy diet

Infections

inflammation

IkB

How do anthocyanins promote endogenous antioxidant

defences and reduce inflammation?


Berry anthocyanins suppress pro-inflammatory

mediators by inhibiting NF-kB nuclear translocation

IL-1, Interleukin 1

TNF-a, Tumor Necrosis Factor a

LPS

bacteria Without

anthocyanins

inflammation

LPS

bacteria With

anthocyanins

lower

inflammation

DIPARTIMENTO DI BIOSCIENZE Lee et al, J Nutr Biochem 25:404, 2014

Omega-6 pathway Linoleic acid LA

Omega-3 pathway a-linolenic acid ALA

Eicosapentaenoic acid EPA

Docosahexanoic acid DHA

Arachidonic acid AA

Our diet can also influence chronic inflammation

through Omega-6 and Omega-3 pathways

Meat, Butter, Eggs

PGE2 LTB4

COX cyclooxygenase

LOX lipooxygenase

PGE3 LTB5

COX LOX

-6 Desaturase

Elongase

-5 Desaturase

Elongase

-5 Desaturase

Oxidation

Pro-inflammatory

eicosanoids Anti-inflammatory

eicosanoids

Safflower, sunflower,

corn, peanut oils Flax, walnuts, canola

OLIVE OIL

15:1

Our diet plays an important role in the balance

between pro-inflammation and anti-inflammation

Pro-inflammatory eicosanoids

Omega-3

Omega-6

Anti-inflammatory eicosanoids

FRIED FOOD SAFFLOWER, SUNFLOWER,

CORN OIL

CHEESE DAIRY, EGGS

RED PROCESSED MEAT SOFT DRINKS VEGETABLES AND FRUITS

SOYBEAN AND

SOY-DERIVED FOOD

BERRIES

WALNUTS AND

FLAXSEEDS

SALMON AND COD


OLIVE OIL

VEGETABLES AND FRUITS

SOYBEAN AND

SOY-DERIVED FOOD

BERRIES

WALNUTS AND

FLAXSEEDS

SALMON AND COD FRIED FOOD SAFFLOWER, SUNFLOWER,

CORN OIL

CHEESE DAIRY, EGGS

RED PROCESSED MEAT SOFT DRINKS

Our diet plays an important role in the balance

between pro-inflammation and anti-inflammation

Omega-3 Omega-6

1:1


Pro-inflammatory eicosanoids Anti-inflammatory eicosanoids

Fish-oil supplementation for 6 months reduces

inflammatory gene expression profiles

EPA+DHA

Omega-3

Blood sample

140 (13.5%)

900

158

Supplementation with fish oil Omega-3

decreased the expression of inflammatory genes,

including NF-kB targets (interleukins),

pro-inflammatory cytokines and eicosanoids

Bouwens et al (2009) Am J Clin Nutr 90:415-24

HA

Omega-6

25 elderly subjects

supplemented with

1.8 g EPA+DHA/day

for 6 months

26 elderly subjects

supplemented with

4 g high-oleic acid/day

for 6 months

Blood sample

Transcriptomic

analysis


Dietary anthocyanins have a fish-like effect

enhancing Omega-3 levels in blood

ACN free diet ACN rich diet

020406080

100120140160

Total ω-3 EPA DHA

marine Omega-3

+17% +41%

+16%

By contrast, Omega-6 levels are unaltered

Toufektsian et al., J Nutr 2011

EPA, eicosapentanoic acid

DHA, docosahexanoic acid


Chronic diseases

Oxidative stress Chronic inflammation

In conclusion, the emerging view is that anthocyanins

act as molecular signals for multiple pathways

Anthocyanins

Nrf2

NfkB RONS Omega-3


Anthocyanin-rich corn as model food

Extracts Seeds Supplements

In vitro cell

experiments

Animal models Clinical trials


Ongoing studies

Epigenetics DNA methylation

Histone acetylation

miRNA

Muscular

distrophy

Inflammatory

bowel disease

Cardiovascular

disease

Intestinal

microbiota


Alzheimer

Epigenetics DNA methylation

Histone acetylation

Cardiovascular

disease


In the frame of the FP7 European Project ATHENA, anthocyanin-rich corn lines are being used to

evaluate:

1) The dose response of anthocyanins in cardiovascular disease

2) Epigenetic mechanisms induced by dietary anthocyanins

3) The potential benefit of anthocyanin-rich supplements to combat excess inflammation

Michel de Lorgeril

Marco Giorgio

Pier Giuseppe Pelicci

Maria Benedetta Donati

Funding agencies

ATHENA partners: Cathie Martin (JIC, UK) coordinator

Hans-Peter Mock (IPK, DE)

M.B. Donati (Neuromed, IT)

Michel De Lorgeril (UJF, FR)

Marco Giorgio (IEO, IT)

Pier Giuseppe Pelicci (IEO, IT)

University of Milan (UNIMI) Dept of BioSciences:

Chiara Tonelli

Katia Petroni

Valentina Calvenzani

Alessandra Marinelli

Monica Fornari

Michela Landoni

Dept of Agr and Env Sciences (UNIMI):

Roberto Pilu, Elena Cassani

Acknowledgments


Collaborations: Graziella Messina (UNIMI, IT)

Michela Matteoli (UNIMI-CNR, IT)

Alberto Mantovani (Humanitas, IT)

Documents

Nutrigenomics: how food influences health how food influences health ... Epigenetics: The ability to ... Fish-oil supplementation for 6 months reduces