Promoting Healthy Aging through Nutrigenomics: The Potential of the Traditional Okinawan Diet

Donald Craig Willcox1, Giovanni Scapagnini2, Bradley J. Willcox11Dept of Geriatric Medicine, University of Hawaii; 2Institute of Neurological

Sciences, Italian National Research Council, Catania, Italy

TODAY`S MENU

Breakfast: Review nutrigenomics in relation to healthy aging

Lunch: Introduce relevant findings from our ongoing research 

projects in Okinawa and Hawaii on healthy aging and  longevity and highlight the connections with nutrigenomics

Dinner: Suggest possible nutritional interventional 

approaches to modulate

gene expression based on 

nutrigenomic properties of the traditional Okinawa Diet

The World`s Oldest PersonThe World`s Oldest Person……EVER !EVER ! 122 years at death122 years at death

Jeanne-Louise Calment of France 1875-1997

Age 20 years

Age 116 years

Projected Increases in Global Population by Age

Source: United Nations Department of Economic and Social Affairs, Population Division. World Population Prospects. The 2004 Revision. New York: United Nations, 2005.

PresenterPresentation NotesThe world's oldest population is the fastest growing

(Age adjusted mortality per 100,000 )Rank Location Life CHD Cancer Stroke All

Expectancy Causes1 Okinawa 81.2 18 97 35 3352 Japan 79.9 22 106 45 3643 Hong Kong 79.1 40 126 40 3934 Sweden 79.0 102 108 38 4358 Italy 78.3 55 135 49 45910 Greece 78.1 55 109 70 44918 USA 76.8 100 132 28 520

WHO Lives the Longest…and WHY ?(who has lowest risk for major killers)

Suzuki et al. Asia Pac J Clin Nutr 2001Suzuki et al. 2001

What is Nutrigenomics?

The study of how different foods can interact with particular  genes to increase or decrease the risk of diseases such as type 

2 diabetes, obesity, heart disease and some cancers

van Ommen B. (2004) Nutrition 20:4-8.

Age‐related Disability & Disease and Lifelong  Accumulation of Cell 

Damage thru Genetic, Environment & Intrinsic (chance) Effects

Accumulation of Cellular Defects

Inflammation

Poor NutritionEnvironmentStress

Anti‐Inflammatory

Healthy 

Nutrition

HealthyLifestyle

Random Molecular Damage

Kirkwood

T. (2005) Cell 120;437‐447

General Research Methods of OCSGeneral Research Methods of OCS

PopulationPopulation--based study based study

(900+ cases 1976(900+ cases 1976--

present)present)

CrossCross--sectional, longitudinal, casesectional, longitudinal, case--

controlcontrol

Age validation Age validation

Geriatric exam with PMH, SH, FH, Geriatric exam with PMH, SH, FH, health habits, anthropometry, ECGhealth habits, anthropometry, ECG

Family pedigreeFamily pedigree

ADLs, IADLs, ADLs, IADLs, psychosocial/cognitive testspsychosocial/cognitive tests

Blood drawBlood draw

The Hawaii Lifespan Studies I and II Defining the Healthy Aging Phenotype (I) NIAR01AG027060 and Genotype (II) 2NIAR01AG027060

POPULATION

8,006 middle‐aged American men of Japanese ancestry from the 

Honolulu Heart Program, followed since 1965

> now over 1200 nonagenarians and centenarians

Hawaii LIFESPAN Study I AIMS

Improve “healthy aging”

phenotypes (better quantify)

Examine (1) environmental and (2) genetic correlates of healthy aging 

and longevity using mainly regression analyses

Focus on insulin‐signaling pathway genes

Hawaii LIFESPAN Study II AIMS

Sequence the FOXO3 gene to find “the”

variant

Understand how the gene reduces mortality Better understand the “longevity mechanism”

Japanese‐

American 

centenarian,  

age 101 years

Interesting “Caloric Restriction (CR) Longevity” Phenotype in Older Okinawans - genetic or environmental?

Less (lifetime) chronic diseaseHigher physical/cognitive function Shorter statureLower BMILower blood sugarLower % T2DMHigher HDLLow cancer

(Willcox et al. Ann NY Acad Sci 2007)

PresenterPresentation NotesVs. Japanese and Americans of similar ageCR　or genetics? Don’t know. Need to compare to other populations? Maybe bec isolated island pop with inbreeding congerate lgene no eveidence form GW study Both genetic and environmental things going on…what is more powerful? Some things changing, still shorter despite env change, could be genetic diffs,

Fontana L et al. Science 2010

Ames and Snell dwarf mice Growth hormone receptor KO mice IGF-1 receptor deficient mice Klotho overexpressing mice Fat Insulin Receptor KO (FIRKO) mice Insulin Receptor Substrate 1 KO mice Brain IRS-2 KO mice Ribosomal S6 protein kinase-1 KO mice p66shc KO mice Type 5 Adenylyl Cyclase KO mice Angiotensin II type 1 receptor KO mice Mice overexpressing catalase targeted to mitochondria

DownregulationInsulin/IGF-1

andmTOR

pathways=

Nutrient –sensingsignalling pathways

Caloric Restriction : Most Powerful AntiCaloric Restriction : Most Powerful Anti‐‐Aging InterventionAging Intervention Calorie Restriction (CR) Reduces Cardiovascular and Cancer MortaCalorie Restriction (CR) Reduces Cardiovascular and Cancer Mortality by 50% in Nonlity by 50% in Non‐‐human Primateshuman Primates

Colman et al. Science 2009

CRAd libitum

P = 0.03HR = 3.0

PresenterPresentation Notes27.6 yrs old = 80 yrs

CR mediated cancer prevention mechanisms

Longo and Fontana. Trends Pharmacol Sci 2010

Long‐term cardiometabolic effects  of CR in humans

Prevention of obesity and type 2  diabetes

Improved lipid profilePowerful anti‐hypertensive and anti‐inflammatory effectsPrevention of atherosclerosisImprovement of left ventricular diastolic 

function (younger hearts) Fontana et al. JAMA 2007Fontana et al. PNAS 2004

Gene Variants in the Insulin Signaling Pathway and Healthy Aging

Genes, Body Size and Longevity

yeast flies miceGene variants that result in lower blood sugar and blood insulin result in long-lived animals with increased resistance to oxidative stress……but what about human beings?

Gene Varient in Insulin Signaling Pathway Strongly Associated with Healthy Aging and Longevity

(Willcox et al. Proc Nat Acad Sci 2008)

Lower prevalence of cancer, CVD, better self- reported health, and higher physical and cognitive function, despite significantly older ages than controls. Greater insulin sensitivity and this was associated with homozygosity for the FOXO3A GG genotype.

“The FOXO Factors”are Master Regulators

An important downstream mechanism whereby FOXO3A may influence human aging is through modification of oxidative stress. In C. elegans, DAF-16 increases the expression of manganese superoxide dismutase (SOD2), which converts superoxide to less damaging hydrogen peroxide and is a potent endogenous protector against free radicals.

Tofu with Fish Lean Meat DishesRice with Vegetables

What do the Okinawan Elders Eat?

Bitter Melon with Tofu Nigana Greens Mozuku Seaweed

Key Features of Traditional Okinawa Diet

1) Low Caloric Density (plant‐based, low fat,  moderate protein from soy, fish, lean meats)2) High Nutrient Density (Vitamins A,C, E, 

potassium, magnesium, folate, and healthy oils)3) Phyto‐nutrient Rich (anti‐oxidants, polyphenols,  flavonoids, carotenoids mostly from green leafy 

and yellow root vegetables)4) Low in Glycemic Load (high quality  carbohydrates from staple sweet potato)5) Anti‐inflammatory (CR, antioxidants, 

polyphenols, flavonoids, omega 3 fatty acids、 curcumins)

Willcox et al (2009) J American College Nutrition 28, (4), 500–516

Okinawan Sweet Potatoes

Kano et al. Antioxidative activity of anthocyanins from purple sweet potato,Ipomoera batatas cultivar Ayamurasaki. Biosci Biotechnol Biochem 69:979–988, 2005.

Sweet Potato: The Staple of the Traditional Okinawa Diet

Willcox et al Journal of American College of Nutrition 2009

USDA: Americans lack sufficient amounts of dietary fiber, calcium, magnesium, potassium, and the antioxidant vitamins A (as carotenoids), C and E.

Gl of Satsuma Imo (Sweet Potato) and Other Potatoes

Willcox et al Journal of American College of Nutrition 2009

http://www.nutritiondata.com/facts/vegetables‐and‐vegetable‐products/2667/2

Okinawa Sweet Potato is highly anti-inflammatory !

The sweet potato truck comes a`calling…..

CURCUMIN : A STRONG ANTI-OXIDANT AND ANTI-INFLAMMATORY

Mice Fed Curcumin and Polyphenols Live Longer Kitani et al (2004) Ann NY Acad Sci 1019:424‐426

Average life span in TC fed mice* was 11.7% longer than in control mice. The 10% longest survival also significantly greater and increase in average life expecancy after 24 months of age was 126%.In mice fed polyphenols average life span increased 6.4% and increase in average life expctancy after 24 months was 72.6%.

* TC: tetrahydrocurcumin * Green tea polyphenols

Soy flavonoids have antioxidant- like effects and hormetic properties and are potent activators of gene expression in FOXO3. Isoflavones, the type of flavonoids most common in soy, also regulate the Akt/FOXO3a/GSK-3beta/AR signaling network in prostate cancer cells. Specifically, they inhibit cell proliferation and foster apoptosis suggestingthat isoflavones might prove useful for the prevention and/or treatment of prostatecancer (Li et al. 2008. Biol Chem. 283, 27707-16.)

Astaxanthin is a xanthophyll carotenoid present in microalgae, fungi, complex plants, seafood (salmon, shrimp, krill). A strong antioxidant with anti-

inflammatory properties with potential as a therapeutic agent in

atherosclerotic cardiovascular disease. There is evidence of a reduction in oxidative stress and inflammation in a small number of clinical studies. This pigment is also commercially produced from the microalga Haematococcus pluvialis, the richest known natural source. (picture is Big Island, Hawaii) Fassett and Coombes Marine Drugs.

2011;

9: 447–465.

Recent evidence suggests that astaxanthin has promise for modulating aging through activation of the insulin signaling pathway and FOXO3 genein particular. Especially, the expression of genes encoding superoxide dismutases and catalases . AX likely protects the cell organelle mitochondria and nucleus of the nematode, resulting in a lifespan extension via Ins/IGF-1 signaling pathway.

Anti-oxidant Activity of Carotenoids

FOXO3a inhibits invasive phenotype of NF639 and rel/CK2-5839 cells

Belguise K et al. Cancer Res 2007;67:5763-5770

©2007 by American Association for Cancer Research

Activation of FOXO3a by the Green Tea PolyphenolEpigallocatechin-3-Gallate ( ECGC) Induces Estrogen Receptor α ExpressionReversing Invasive Phenotype of Breast Cancer Cells

PresenterPresentation NotesFOXO3a inhibits invasive phenotype of NF639 and rel/CK2-5839 cells. A, NF639 cells were infected with a retrovirus control (pBabe) or retrovirus expressing A3 FOXO3a (A3). Whole cell protein extracts were subjected to immunoblotting for FOXO3a, E-cadherin (E-cad), Snail, and β-actin, which showed equal protein loading. B, NF639 cells were infected with a retrovirus control (pBabe) or retrovirus expressing A3 FOXO3a and grown either in Matrigel or on plastic. Top, after 6 d, the colonies in Matrigel were photographed at a magnification of 50×. Representative fields are shown. Bottom, NF639 cells infected with control pBabe or pBabe expressing A3 FOXO3a were seeded at a density of 20,000 cells per well in 24-well cell culture dishes. After 3 d, both cultures reached confluence. C, NF639 cells were infected with pBabe-puro-A3 FOXO3a (A3) or pBabe-puro (pBabe) retrovirus. After 7 d of selection with puromycin, 5 × 105 cells were subjected to a migration assay for 6 h. OD410 nm values correspond to cells that migrated to the lower side of the filter and represent the mean of three determinations. D, rel/CK2-5839 cells were infected with pBabe-puro-A3 FOXO3a or pBabe-puro retrovirus. After 7 d of selection with puromycin, 5 × 105 cells were subjected to a migration assay for 6 h. OD410 nm values correspond to cells that migrated to the lower side of the filter and represent the mean of three determinations. Inset, whole cell protein extracts prepared from rel/CK2-5839 cells infected with control pBabe or pBabe expressing A3 FOXO3a were subjected to immunoblotting for Snail and β-actin.

Okinawa Traditional Dietary Intervention  Study in Americans 

Vacuum Packed  Bento

Okinawa Diet Intervention Achieves “DASH*-like” Blood Pressure Reductions in Americans

70

80

90

100

110

120

130

0 2 3

M onth

Blood pressure(m

mHg)

S B P (A group)

S B P (B group)

D B P (A group)

D B P (B group)

A group intervention

B group intervention

1. SBP reduced 2.6 mm Hg (95% CI -4.3, -1.2. 2. DBP reduced 2.1mm Hg (95%CI -3.1 -1.0)

and 0.3mmHg (95%CI -2.1 0.6).

3. 24h-urinary sodium and body weight reduced (between-group differences ranged from p=0.032 to 0.0002).

Todoriki et al J Hypertension 2008

*Dietary Approaches to Stop Hypertension (DASH:

Most common physician prescribed diet to lower high blood pressure

in the U.S.

Try it !

Even in Very Late Life

(Nonagenarians) Survival is Longest in those with 

Healthy Diet and

Good Genes (participants from Honolulu Heart Program) 

HDI = Healthy Diet Index (ate healthy diet); FOXO3+ indicates prevalence of protective allele

Conclusions

Chronic low grade inflammation may be a major 

determinant of ageing rate and longevity

Recent research has uncovered important “longevity 

genes”

and pathways that regulate the aging process (ie.  Foxo)

Nutritional approaches such as flavonoids, polyphenols, 

omega 3 fatty acids, and calorie restriction are promising  and further research is needed in dietary, cell‐based and 

pharmacological strategies for healthy aging

The Okinawa diet contains many such compounds with 

potentially powerful nutrigenomic

effects and research is  underway in order to isolate the most promising 

candidates (eg. Sweet potato extracts, curcumin, soy  flavonoids, sanpin

and green tea, etc.) that will up‐

regulate the most promising target genes.

UshiUshi--sansan 102 Years Young and 102 Years Young and Still Still DigginDiggin’’ LifeLife

Mahalo

!

Domo Arigato! Domo Arigato!

Support & Financial Disclosure

The investigators retained full independence in the conduct of this 

research and acknowledge the support from:

The John A. Hartford Center of Excellence in Geriatrics, Dep’t of 

Geriatric Medicine, John A. Burns School of Medicine, University

of 

Hawaii

Kuakini

Medical Center

US National Institute on Aging

US National Heart, Lung, and Blood Institute

Japan Society for the Promotion of Science  

Special thanks to Dr. Makoto Suzuki (PI of Okinawa Centenarian Study) 

and Dr. Hidemi

Todoriki

(PI of Chample

Study). For more information 

on the Okinawa centenarian Study see www.okicent.org

Dr. Donald CraigWillcox

is Head of the Scientific Advisory Board for 

Koruchem

Therapeutix, an evidence‐based pharmaceutical and 

nutra/cosmaceutical

company 

(www.koruchemtl.com) 

http://www.koruchemtl.com/

Slide Number 1Today`s Menu Slide Number 3The World`s Oldest Person…EVER !� 122 years at death Slide Number 5Slide Number 6Slide Number 7 What is Nutrigenomics?Slide Number 9 Age-related Disability & Disease and Lifelong Accumulation of Cell � Damage thru Genetic, Environment & Intrinsic (chance) Effects�Slide Number 11Slide Number 12�The Hawaii Lifespan Studies I and II�Defining the Healthy Aging Phenotype (I) NIAR01AG027060 and Genotype (II) 2NIAR01AG027060Interesting “Caloric Restriction (CR) Longevity” Phenotype in Older Okinawans - genetic or environmental? Slide Number 15Slide Number 16Slide Number 17Slide Number 18Caloric Restriction : Most Powerful Anti-Aging Intervention�Calorie Restriction (CR) Reduces Cardiovascular and Cancer Mortality by 50% in Non-human Primates CR mediated cancer prevention mechanismsLong-term cardiometabolic effects �of CR in humansSlide Number 22Slide Number 23Slide Number 24 “The FOXO Factors”are Master Regulators�Slide Number 26Slide Number 27Key Features of Traditional Okinawa DietSlide Number 29 Sweet Potato: The Staple of the Traditional Okinawa Diet Gl of Satsuma Imo (Sweet Potato) and Other PotatoesSlide Number 32Slide Number 33Slide Number 34Mice Fed Curcumin and Polyphenols Live Longer�Kitani et al (2004) Ann NY Acad Sci 1019:424-426Slide Number 36Slide Number 37Slide Number 38Slide Number 39Slide Number 40Slide Number 41Okinawa Traditional Dietary Intervention Study in Americans Vacuum Packed Bento� Okinawa Diet Intervention Achieves “DASH*-like” � Blood Pressure Reductions in Americans Slide Number 45� ���� Even in Very Late Life (Nonagenarians) Survival is Longest in those with � Healthy Diet and Good Genes (participants from Honolulu Heart Program) � ConclusionsSlide Number 48 Support & Financial Disclosure�