Nutrigenomics:
how food influences health
Dr. Katia Petroni
Dipartimento di Bioscienze
Università degli Studi di Milano
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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
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
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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
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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
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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
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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
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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
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DNA methylation
Methylating the cytosine of a CpG
motif results in gene silencing and
silent heterochromatin
Nutrients
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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
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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
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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!
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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
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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
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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
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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
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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
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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?
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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)
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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?
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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
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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
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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
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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
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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
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Anthocyanin-rich corn as model food
Extracts Seeds Supplements
In vitro cell
experiments
Animal models Clinical trials
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Ongoing studies
Epigenetics DNA methylation
Histone acetylation
miRNA
Muscular
distrophy
Inflammatory
bowel disease
Cardiovascular
disease
Intestinal
microbiota
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Alzheimer
Epigenetics DNA methylation
Histone acetylation
Cardiovascular
disease
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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
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Collaborations: Graziella Messina (UNIMI, IT)
Michela Matteoli (UNIMI-CNR, IT)
Alberto Mantovani (Humanitas, IT)