Transcript Document

Genomics in the Science of
Nutrition
John Milner
Nutritional Science Research Group
Division Cancer Prevention,
National Cancer Institute
[email protected]
While there are opportunities for expanded use of foods, and their
components, to achieve one’s genetic potential, increase productivity
and reduce the risk of disease.
Hippocrates Proclaimed
almost 2500 years ago:
Let thy food be thy medicine
and thy medicine be thy food
Worldwide Leading Causes of Mortality
(2002)
Aged 60>
Aged 15-59
Death
Rank
Cause
1
HIV/AIDS9
(000)
227
Death
Rank
Cause
1
Ischemic heart disease
(000)
5825
2
Ischemic Heart Disease
1332
2
Cerebrovascular disease
4689
3
Tuberculosis
1036
3
Chron obste pulmon disease
2399
4
Road traffic injuries
814
4
Lower respiratory infections
1396
5
Cerebrovascular disease
783
5
Trachea, bronchus, lung cancer 928
6
Self-inflicted injuries
672
6
Diabetes mellitus
754
7
Violence
473
7
Hypertensive heart disease
735
8
Cirrhosis of the liver
382
8
Stomach cancer
605
9
Lower respiratory infections
352
9
Tuberculosis
495
10
Chron obstr pulmon disease
343
10
Colon and rectum cancers
477
Charts, Maps, Tables, The World Health Report, WHO 2003
5 of the 10
Leading Causes of
Death of
Americans Relate
to Dietary Habits
Age-Adjusted Incidence Cancer Rates in Males by Region
Australia/New Zealand
Northern America
Western Europe
Eastern Europe
Southern Europe
Northern Europe
Southern Africa
Eastern Asia
South America
Polynesia
Caribbean
Central America
Eastern Africa
Micronesia
Western Asia
Melanesia
Middle Africa
South-Eastern Asia
Northern Africa
South Central Asia
358.58
357.43
318.69
290.01
275.39
263.37
217.53
205.33
201.35
200.7
187.91
178.54
177.68
Ferlay et al. (2001)
ARC CancerBase No. 5.
Lyon, IARCPress
175.45
151.13
149.84
141.82
131.13
124.46
106.61
Rate/100,000 (adjusted to world standard population)
WHO South-East Asia Report (Oct 2005) Suggests:
At least 80% of premature heart disease, stroke and
type 2 diabetes, and 40% of cancer could be
prevented through a healthy diet, regular physical
exercise and avoiding tobacco products.
In the South-East Asia Region, a 2% annual
reduction in deaths due to chronic disease could
save over 8 million lives in the next 10 years.
In India, a similar reduction would also result in an
economic gain of USD 15 billion over the next 10
years.
Public Health Approach
U.S. Nutrition Industry
($billion in consumer sales in 2005 )
Supplements
2004
20.4
2005 % Change
21.18
3.8
Natural/Organic Foods
18.38
20.70
12.7
Functional Foods
24.46
26.66
9.0
5.47
5.94
8.5
68.71
74.48
8.4
N & O Personal Care
Total
Nutrition Business Journal, interim research based on a variety of
sources as of April 28, 2006
Much Confusion Exist
It Is Illogical to Assume
All Individuals are
Equally Responsive to
Drugs, Foods or Food
Components!!
Lots of Variability in Response Across Studies
Asian
Western
Lee ‘92 (total soy protein)
p < 0.001 Premenopausal
NS Postmenopausal
Hirose ‘95 (bean curd, miso)
Yuan ‘95 (tofu, soymilk)
NS Premenopausal
NS Postmenopausal
NS p = 0.44–0.79 Shanghai, Tianjin
Wu ‘96 (tofu)
p < 0.01 Premenopausal
p < 0.05 Postmenopausal
Key ’99 (soy)
Tofu
Miso
Zheng ’99 (urinary isoflavonoids)
Dai ‘01 (soy)
NS All Breast Cancer
S Just ER+/PR+
Wu ’02 (soy)
Yamamoto ’03 (isoflavonoid consumption)
Premenopausal
Postmenopausal
Wu ’04 (soy)
Ingram ‘97 (urinary isoflavones)
NS Diadzein
p = 0.009 Equol
Witte ’97 (soy)
den Tonkelaar ‘01 (urinary phytoestrogens)
NS Postmenopausal
Horn-Ross ’01 (phytoestrogen intake)
Keinan-Boker ‘02 (food content)
NS Isoflavones
S Lignans
Linseisen ’04 (isoflavone intake)
daidzein and genistein
.1
.3
.6
.6
1.3
.6
.8
.7
2.6
1.0
1.0 1.3
.61
.6
.78
1.47 .87
.68
1.12 .65
.18
2.37
.46
.25 .44
.66
1.07
.95
.78
.4
.6
1.2
.22
.48
1.1
.25
.47
.90
.36 .53 .78
.17
.47
.1
1.33
.27
.2
.69
.5
1.1
.46
.83
.79
.79
.34
.36
1.0
1.08
.58 .98
.57
1.3
.83
1.3
1.59
The “Omics” of Nutrition
Bioactive
Food
Component
N
u
t
r
i
g
e
n
o
m
i
c
s
Nutrigenetics
DNA
Nutritional
Epigenetics
RNA
Nutritional
Transcriptomics
Phenotype
Protein
Proteomics
Metabolomics
Metabolite
Nutritional Preemption
(A Strategy for Health Promotion)
Concept that bioactive food
components can be introduced
at points of initiation &
progression for pathway leading
to an unhealthy or lethal
phenotype
Appeal for Nutrigenomics
• Individualized/targeted
•
•
•
•
approach
Recognizes cultural
differences
Hi-technological Approach
New Market Niches
Suitable for capital
investments from food and
pharmaceutical companies
Skeptics
•
•
•
Diet and public health
already too complex
Only for rich can afford,
create more inequalities
Ethical minefield
• Essential
Nutrients- Ca,
Zn, Se, Folate, C, E
• Non-Essential
PhytochemicalsCarotenoids, Flavonoids,
Indoles, Isothiocyanates,
Allyl Sulfur
Zoochemicals - Conjugated
linoleic acid, n-3 fatty acids
Fungochemicals - Several
compounds in mushrooms
Bacteriochemical - Those
formed from food
fermentations and those
resulting from intestinal
flora
Complexity is Illustrated by the Vast Number of Foods and Components which
can modify the cancer process
Variation in Glucosinolate Among
Cruciferous vegetables (µmol/g DryWeight)
12
progoitrin
glucoraphanin
glucobrassicin
sinigrin
10
8
6
4
2
0
Broccoli
Brussels
Cabbage Cauliflower
Kushad et al, J Agric Food Chem. 47(4):1541, 1999
Kale
Types of Bioactive Compounds May Vary
with New Varieties
Tomato Varieties
Intake of all Tomatoes and Tomato Products
(g/day, log transformed)
Individual Consumption of All Tomatoes and Tomato
Products and Serum Lycopene Levels
(EPIC Cross-sectional Study in 3000 subjects)
6.0
Corr = 0.23
5.0
4.0
3.0
2.0
1.0
0.0
0.0
1.0
2.0
3.0
4.0
5.0
Mean Serum Lycopene (microg/dl, log transformed)
Jenab et al. J. Nutr. 135:2032, 2005
6.0
Daidzein in Soy Converted to Equol in
Some but not All Individuals!
30-50% of Population
80-90% of Population
Intestinal Bacteria
F Rafii et al,
Arch
Microbiol
180: 13, 2003
Tissue Polyphenols (ng/g tissue)
Plasma and Tissue Concentrations
Are Not Always Identical
1000
EGC
EC
EGCG
800
600
400
200
0
Plasma
Colon
Kim et al., Nutrition and Cancer 37: 41-48, 2000
Bladder
Lung
Bronchodilator Response to Albuterol
% Responding
60
50
40
30
20
10
0
Gly-Gly
Arg-Gly
Arg-Arg
b-2 andrenergic Receptor Genotype (codon 27)
Drazen et al , Int Arch Allergy Immunol 124: 183, 2001
Dietary Calcium, VDR FokI
Genotype and Colon Cancer Risk
OR for Colon Cancer
3
*
2.5
Dietary Calcium
< 388 mg/day
*
2
>388 mg/day
1.5
P for trend=0.004
1
0.5
0
FF
Ff
ff
VDR Genotype
Wong et al. Carcinogenesis, 24: 1091-1095, 2003
Change in Diastolic Blood Pressure in Response
to Fiber Based on AGT Gene Codon 235
2
1
D DBP (mm Hg)
0
-1
-2
Insoluble
-3
Soluble
-4
-5
-6
T/T
T/M
Hegele, et al. Nutr. Res. 17: 1229, 1997
M/M
Inflammation Often Linked to Several Diseases:
(Fish Oil Response Depends on TNF-α Polymorphism)
12
TNF-α Production (µg/l)
Control
10
Fish Oil
8
6
**
4
2
0
G/G
G/A
TNF-α Polymorphism at -308
Grimble et al. Am J Clin Nutr 76:454-9, 2002.
Influence of Caffeine on Bone Mass May Depend on Genes
6
< 300 mg
Caffeine
> 300 mg
0
-6
-12
TT
Tt
tt
TT
Tt
Vitamin D Receptor Genotype
Rapuri et al. Am J Clin Nutr 2001 Nov;74(5):694-700
tt
Not Knowing Genomics Can Cause Misinterpretations
p21+/+ AIN-76A Diet
Percent survival
100
90
p21+/- AIN-76A Diet
80
p21+/+ Western Diet
70
p21-/- AIN-76A Diet
p21+/- Western Diet
60
50
40
30
p21-/- Western Diet
20
10
0
0
4
8
12
Yang et al, Cancer Res. 61, 565, 2001
16
20
24
Weeks
28
32
36
Nutrigenomic Testing
Promises versus Reality!
• Commercial Nutrition-Gene Test
– Genelex Sciona 19 genes including
MTHFR $395
– Gene Care CVD nutritional genetic test
(South Africa) MTHFR (Hcyst),
apoA1 (HDL) +9 others $400
• Exceeding complex area since about
30, 000 Genes, 5-8 Million SNPs
% of Animals with Tumors
Transgenic and Knockout Models
Key to Identifying Sites of Action of
Food Components
100
Prostate Tumors in Lady
mice.
80
Antioxidants= vitamin E,
selenium and lycopene
60
40
20
*
*
0
Control
High Fat
Control + Anti
Venkateswaran et al., Cancer Research 64: 5891-5896, 2004
High Fat + Anti
Diet May Influence Genetic and Epigenetics!
Epigenetics: The study of heritable changes in gene expression
that occur without a change in DNA sequence.
– DNA methylation- CpG islands
– Histone posttranslational modifications:
Acetylation of lysines
Methylation of lysines and arginines
Phosphorylations of serines and threonines
ADP-ribosylation of glutamic acids
Ubiquitination of lysine residues
Sumolyation of lysine residues
Biotinylation of lysines
– Recently, in human cells small-interfering
RNAs (siRNAs) have been shown to mediate
transcriptional gene silencing (Morris KV, Cell Mol.
Life Sci. 62:3057-3066, 2005).
LTR Hypomethylated
When to
Intervene??
LTR Hypermethylated
Maternal
Supplements
with
Yellow Mouse
High risk cancer, diabetes,
obesity & reduced lifespan
zinc
methionine
betaine
choline,
folate
B12
Cooney et al. J Nutr 132:2393S (2002)
Agouti Mouse
Lower risk of cancer, diabetes,
obesity and prolonged life
Genistein Can Also Influence Agouti Phenotype
Viable yellow Agouti (Avy) Locus
40
Control Diet
Control Diet
100
Genistein Diet
Genistein Diet
p = 0.007
30
Pseudo-agouti
76%
Site Specific Methylattion (%)
Avy Offspring (% of Total)
75
20
10
Heavily Mottled
57%
50
Mottled
32%
25
Slightly Mottled
15%
Yellow
8%
0
0
4
Yellow
Slightly
Mottled
Mottled
Heavily
Mottled
Pseudoagouti
5
6
7
8
9
4
5
6
7
CpG Site Number
Dolinoy et al. Envir. Health Perspect. 114: ePublished, 2006
8
9
Histones Can Be Regulated by Isothiocyanates,
Allyl Sulfur, Genistein and Butyrate
HDAC
Co-repressor
complex
Acetylated histones H3 and H4
associated with P21 and Bax promoters
HDAC Inhibition
Molecular Target??
Transcription of P21 and Bax mRNA
HDAC
Co-repressor
complex
acetyl groups added
p21 and Bax protein levels increased
Cell cycle arrest
Caspase activation
APOPTOSIS
Transcriptomic Studies Are Providing Clues About
Molecular Targets for Specific Food Components
Corn Oil
Fish Oil
Olive Oil
Davidson et al., Cancer Res. 64: 6797-6804, 2004
A Molecular Approach to Medicine
Recent News Release suggests:
Herceptin is a Novel Pioneering Drug for
Personalized Medicine Approach Based
on Pharmacogenomics to block Her2-neu
expression.
Evidence Has Existed for
Years that:
EGCG from Green Tea, Oleic Acid from
Olive Oil, and n-3 fatty acids from Fish Oil
and Apigenin from parsley, thyme, and
peppermint can significantly influence
HER2neu expression!
So why are we not being more proactive in
showcasing dietary effects?
Must Focus on Processes Which Are Modified by
Single or Multiple Foods/Components
Cellular
Division
Compound
Activation
DNA
Repair
Apoptosis
Digestion
Bioactive
Food
Components
Differentiation
“Credentialing”
Cellular
Energetics
Hormonal
Homeostasis
Inflammation
Immunity
Pentameric procyanidin (100ug/ml) from Theobroma cacao
Can Inhibit Human Breast Cancer Cells
Ramljak et al. (2005) Mol. Cancer Ther 4:537-546
Likely Molecular Targets
For Isothiocyanates, Allyl Sulfur, Selenium, etc.
Bioactive Component
GST
QR
GST
QR
Proteomics and Metabolomic Technologies and
the Response to Bioactive Food Components
Mitchell et al. (2005) Proteomics 5:
2238 Identified 2 peaks that reflect
with 76% accuracy broccoli intake
Wenzel et al. (2004) Proteomics
4:2160 Various proteins modified by
quercetin
Yang et al (2004) Obesity Res 12:1179
Exercise and quantitative proteomics
Mayr et al. (2005) Arterioscler
Thromb Vasc Biol. 25:2135.
Metabolomics demonstrate gene
interactions with inflammation,
oxidative stress, and energy
metabolism.
Metabolomic Analysis of the Biochemical
Effects of Dietary Isoflavones
Pre-Soy
Post-Soy
Solanky et al., analytical Biochemistry 323: 197-204, 2003
Another Questions Must Be “What is the Amount
Needed to Bring About a Desired Outcome?”
Typical Intakes??
Toxicity
Response
Cell Cycle
Inhibition
Apoptosis
Immune
Enhancement
Antioxidant
Differentiation
Carcinogen
Metabolism
Nutritional
Supranutritional
Toxic
Exposure
Modified from Combs and Gray, Parmacol. Ther. 79: 179-192, 1998.
Components are “complex
mixtures” - act synergistically
Combination of Dietary Components
Dietary Component
% Reduction
 Wine (150 ml/day)
32%
 Fish (114 g 4x/week)
14%
 Dark Chocolate (100 g/day)
21%
 Fruits and Vegetables
21%
(400 g/day)
 Garlic (2.7 g/day)
25%
 Almonds (68 g/day)
12%
76% decreased risk of CVD
Franco et al. BMJ 329:1447-1450, 2004
Soy Phytochemicals and Green Tea Inhibit
Human Mammary Tumors in Mice
Zhou et al, Int. J. Cancer 108: 8-14, 2004
Is it ever too later to intervene?
When should one stop
looking for a preemptive response?
Kaplan-Meier Estimates of the Cumulative Hazard for
Invasive Breast Cancer
Copyright restrictions may apply.
Prentice, R. L. et al. JAMA 2006;295:629-642.
Women’s Intervention Nutrition Study (WINS)
RELAPSE EVENTS (ER NEGATIVE)
Diet
28/205
PATIENTS (%)
30
Control
59/273
HR, 95% CI
0.58,0.37-0.91
p-value
0.018
4
6
25
Control
Diet
20
15
10
5
0
0
1
2
3
5
Follow-up time (Years)
P-value from adjusted Cox proportional hazard model
Source: Chlebowski RT et al. Presentation. ASCO 2005.
7
8
Does the Biological Response in Pathologic Conditions
Reflect Normal Response ??
Diet
Biological Response
Physiological
Function
???
Pathological
Function
Functional/Phenotypic Consequences
Dual Association of b-carotene with risk of
tobacco-related cancers in a cohort of
French women.
b-carotene intake was inversely associated with
risk of tobacco-related cancers among
nonsmokers with a statistically significant dosedependent relationship, whereas high betacarotene intake was directly associated with risk
among smokers.
Touvier et al. J Natl Cancer Inst. 2005;97(18):1338-44.
1a Hydroxylase Activity in Normal (N) Benign
Prostatic Hyperplasia (BPH), Prostate Cancer Tissue
(PC) and Human Tumor Cells.
Schwartz (2005) Seminars in Dialysis 18:276.
When I knew all of life’s answers,
they changed all the questions!
How Will We Optimize Benefits and Minimize Harm?
New Research Paradigm-New Hope
Discovery
Development
Delivery
Discovery of relevant mechanisms
Development of interventions for better detection,
diagnosis, treatment, and prevention
Delivery to all in need
Three Types Biomakers Needed To Identify
Responders (Both Positive and Negative)
Absorbed
Dose
Dietary
Exposur
e
Health Effects
+ and -
Inactive Metabolite
Biologically
Effective Dose
Susceptibility
Factors
Altered
Structure/
Structure
Function
Molecular
Target
Early
Biologic
Effect
The Future of Nutrigenomics
Rest With The:
•Identify and validate nutrigenetic,
nutritional epigenetic & transcriptomic,
proteomic and metabolomic biomarkers of
effect and susceptibility
•Effective communicate about “omics”
information to the health care community
and consumers
•Ability to work within a responsible
bioethical framework
Develop Partnerships Which Build on Strengths
Occurring In A Borderless Society
Foster Triple Helix- Academia, Industry and Government