Rafael de CaboAging, Interventions and Metabolism Section

Laboratory of Experimental GerontologyNational Institute on Aging

National Institutes of Health

Nutrition and Aging

What is Aging? What are the mechanisms behind aging?

Can we manipulate the aging process?

Metabolic (Nutrition)

Genetic

Environment

Mean Lifespan

Maximum Lifespan

Heart DiseaseCancerStrokeEmphysemaPneumoniaDiabetesAccidentsKidney DiseaseAlzheimer’s

15-2425-34

35-4445-54

55-6465-74

75-8484+

AGE GROUP

Dea

ths

per 1

00,0

00 p

er y

ear 100000

10000

1000

100

10

1

0

Aging is the major risk factor for ALLchronic diseases

Heart DiseaseCancerStrokeEmphysemaPneumoniaDiabetesAccidentsKidney DiseaseAlzheimer’s

15-2425-34

35-4445-54

55-6465-74

75-8485-94

AGE GROUP

Dea

ths

per 1

00,0

00 p

er y

ear 100000

10000

1000

100

10

1

0

95-104105-114

0

20

40

60

80

100

0 6 12 18 24 30 36 42

% S

urvi

val

ΔHea

lthsp

an

ΔLifespan

% O

ptim

al H

ealth

Manipulating Lifespan/Healthspan

Time (weeks)50 60 70 80 90 100 110 120 130 140 150 160

Surv

ival

pro

port

ion

0.0

0.2

0.4

0.6

0.8

1.0 CR

AL

HF

Lifespan Manipulation C57Bl6/NIA

Aging Intervention Program; Working Hypothesis

Resveratrol

• Resveratrol is a phytoalexinproduced in response to stress (e.g. infection, heat, UV).

• Found in wine (0.2-5.8 mg/L wine).

• Cancer protection, inflammation and antioxidant.

• Resveratrol activates SIRT1 in vitro.

• It is similar to caloric restriction in that there is much debate about its mechanism (s).

OH

OH

HO

3572, today

Aging Intervention Mouse Program

11 Groups, n≥50 mice/group

HealthspanLifespan

Food IntakeBody Temperature

BehaviorSerum/UrineAnalysis

Glucose Homeostasis

Tissue Analysis

Metabolic assessment

Resveratrol 100 and 400 mg/kg

NMRBody Comp

Immune/Stress test

1 year old male C57Bl6/J

C57BL/6J• Males• Aged (1y)• n ≥ 50/group• Microchiped

(ID+Temptransponders)

Lifespan data

Time (weeks)50 60 70 80 90 100 110 120 130 140 150 160

Surv

ival

pro

port

ion

0.0

0.2

0.4

0.6

0.8

1.0

Standard diet control

SD high resv

EOD low resv

High calorie control

HC high resv

Health and Lifespan Summary

• Protection against fatty liver induced by HF• Increased insulin sensitivity• Gene expression pattern similar to standard diet

for HF mice and to CR for SD mice• Increased performance on a rotarod• Mean and maximum lifespan extension in 3 groups

(HF+HR, HF+LR and EOD+LR)• Reduction in cataracts• Improve bone health parameters• Reduced damage/stress in cardiovascular system

Resveratrol Non-Human Primate Work-in-progress!

Will Resveratrol Improve CardiovascularFunction In Monkeys on a High Fat Diet?

Julie Mattison and Kevin Pearson

Nonhuman primates

• Resveratrol study• 24 Male rhesus monkeys (Macaca mulatta)• age range: 7‐13 years (full adult)• body mass range: 7‐20 kg• 24 month study

24 monkeysStandard Diet (SD)

Group A Group B Group C10 HFS + Resv 10 HFS 4 SD control

Baseline Collection

Time 0

3 months

6 months

9 months

12 months

15 months

18 months

21 months

24 months

42% kcal fat27% kcal sucrosePlacebo twice daily

42% kcal fat27% kcal sucrose40 mg RESV twice daily

Groups and Timeline

13% kcal fat<5% sucrosePlacebo twicedaily

Sacrifice and tissue collectionSacrifice and tissue collection

Increase RESV dose to 240 mgtwice daily

Body Weight & CompositionBody Weight

10

11

12

13

14

15

16

17

18

19

20

Baseli

ne3 M

onth 40

mg6 M

onth 40

mg9 M

onth 40

mg12

Month

40mg

3 month 24

0mg

6 month 24

09 m

onth 240

12 m

onth 24

0

Avg

Wei

ght (

kg) Control HFS HFS + Resveratrol

0

1

2

3

4

5

6

7

8

Baseline 1 yr 40 1 yr 240

HFS

RESV

LEA

N:F

AT

Carotid (A)

Femoral (B)

D

Carotid

Femoral

Pulse Wave Velocity

ΔT

Pulse Wave Velocity

*p<0.01#p<0.05

Pulse Wave Velocity

700900

1100130015001700190021002300

0 6 12 15

PW

V (c

m/s

ec)

HFSResv

18 24

40 mg twice/day 240 mg twice/day

* * *

#

Parametric Analysis of Gene Set Enrichment (PAGE)

http://www.broad.mit.edu/gsea/msigdb/msigdb_index.html

Uses a priori defined groupings of genes into functional pathways

(Zsc

ore)

AF_

AC

(Zsc

ore)

AR

_AF

(Zsc

ore)

AF_

AC

(Zsc

ore)

AR

_AF

(Zsc

ore)

VF_

VC

(Zsc

ore)

VR_V

F

(Zsc

ore)

VF_

VC

(Zsc

ore)

VR_V

F

(Zsc

ore)

FHF2

_FC

2

(Zsc

ore)

FRS2

_FH

F2

(Zsc

ore)

FHF2

_FC

2

(Zsc

ore)

FRS2

_FH

F2

(Zsc

ore)

MH

F2_M

C2

(Zsc

ore)

MR

S2_

MH

F2

(Zsc

ore)

MH

F2_M

C2

(Zsc

ore)

MR

S2_

MH

F2

Aorta Ventricle Fat Muscle

HF:

SD

HFR

:HF

HF:

SD

HFR

:HF

HF:

SD

HFR

:HF

HF:

SD

HFR

:HF

Pathways up in HC

Pathways down in HC

Resveratrol opposes the transcriptional effects of a high calorie diet in Rhesus Monkeys, all 24 monkeys were arrayed

Microarray analysisParametric analysis of gene set enrichment

Uses a priori defined groupings of genes into functional pathways

Heart MuscleMonkey vs. Mice, Genes Reversed by Resveratrol

Fat

NDUFA5NDUFB8

MYL6DNAJB6

IL6STCYP1B1

IGF1CRYAB

GSTM1CEBPB

GPX4ATP5B

EIF4A1ATF3

EEF1GARF3

EEF1B2APOD

Preliminary Conclusions/Future Directions

• Resveratrol had a strong cardiovascular protective effect in NHP

• Improvement on PWV, an age related clinical marker of atherosclerosis

• Reversal of the gene transcription profiles in multiple tissues

• Overlap of genes/pathways in both species

• Studying mechanisms behind resveratrol’s effects• Human Intervention study with Luigi Ferrucci at CRB

Acknowledgements

• Office of the Scientific Director• Laboratory of Experimental

Gerontology• Kevin Pearson• Kaitlyn Lewis• Nate Price• Julie Mattison• Dawn Boyer

• Research Resources Branch• Comparative Medicine Section• Gene Expression and Genomics Unit

• Laboratory of Cardiovascular Science• Laboratory of Clinical Investigation

• Office of Dietary Supplements• David Sinclair (Harvard)• Joe Baur (U. Penn)• Norm Wolf (Washington)• David Le Couteur (U. Sydney)• Placido Navas (U. Sevilla)• Zoltan Ungvari (NYMC)

“See, the problem with doing things to prolong your life is that all the extra years come at the end, when you’re old.”