David Sinclair, Ph.D.

Departments of Pathology and Genetics

Harvard Medical SchoolBoston

Disclosure statement: DAS is a consultant to Sirtris and Genocea.

EnvironmentalCues

Causes of Disease& Aging

Longevity RegulationPathways

Small Molecules

ERCs

SIR2 SIR2homologs

LOWGLUCOSE

LOWAA

PNC1

AGING

TOR,Msn2/4

Anderson et al., Nature (2004)Lamming et al., Science (2005)Medvedik et al., PLoS Biol. (2007)

NAD+

NAM

HEATSHOCK

2% glucose 0.5% glucose 0.1% glucose low amino acids low nitrogen

Anderson et al., Nature (2003)

yeast Sir2

SIRT1

SIRT2

SIRT3

SIRT4SIRT5

core domain

SIRT6

SIRT7

Humans possess 7 NAD+-dependent deacetylasesa.k.a. “sirtuins”

humans

Save the date: Sirtuin session tomorrow 5 PM

Sirtuins

NAM

NANAMN

NaAD+

SIRTsPARPs

NMN

Yeast,Flies,

nematodesProteobacteria,

Mammals

Nampt(PBEF)

Pnc1

The NAD+ salvage pathway

NAD+ NAD+ NAM

ANT

CypDAc 166

PTP

NAD+

Mitochondrial matrix

SIRT3

ANT

CypD

Ac166

PTP

Mitochondrial matrix

SIRT3

ANT

CypD

Ac166

PTP

Mitochondrial matrix

SIRT3

ANT

DepolarizationReduced ATPMitochondrial destruction

Leakage of solutes out of mitochondria

Neurodegeneration, Cardiac hypertrophy

CypD

Ac166

PTP

Mitochondrial matrix

SIRT3

Haigis and Sinclair, Annual Reviews, 2010

Circadian clock

DietExercise

Quercetin (4.6x)

Fisetin (6.6x)

Piceatannol (9.0x)

Butein (8.5x)Resveratrol (13.0x)

Isoliquiritigenin (7.6x)

Representatives from three classes of plant-derived STACs

Stilbenes Chalcones Flavones

Howitz et al., Nature, Aug (2003)

Gruber et al., 2007 Valenzano et al., 2006Jarolim et al., 2004

Calorie Restriction (EOD)

High CalorieDiet (HC)

Lean Diet(SD)

ActivatorActivator

Rafa deCabo

Standarddiet

High calorie diet+ resveratrol

15 month old mice

High caloriediet

**photo by Doug Hansen

Oil Red O fat stain

LIVER

Fixed whole liver

H&E

Standard diet(SD)

High calorie(HC)

High calorie+ resv (HCR)

Lagouge et al., Cell (2006)

Baur et al., Nature (2006)

Last mouse Resveratrol fed every other day

36 months of age

Howitz and Sinclair, Cell (2008)

Generation of inducible SIRT1‐knockout mice

E1 E2 E3 E5 E6 E7 E8 E9E4

NEO

Cre‐ERT2

E1 E2 E3 E5 E6 E7 E8 E9

E1 E2 E3 E5 E6 E7 E8 E9

E4

Cre‐ERT2(Cytoplasmic)

Cre‐ERT2(Nuclear)

Tamoxifen

Cre‐ERT2

Cre‐ERT2

E4

0

10000

20000

30000

40000

50000

60000

70000

StD WT StD T1KO HFD WT HFD T1KO

HFD Low RSV WT

HFD Low RSV

T1KO

HFD High RSV WT

HFD High RSV

T1KO

Rhod

amine 123 Fluo

rescen

ce(RFU

s)

WT KO WT KO WT KO WT KOStandard

Diet High Fat

DietHigh Fat

Diet + resveratrol

Mitochondrial membrane potential

Low dose High Dose

Low dose = 100 mg/kg/dHigh dose = 400 mg/kg/d

0

0.5

1

1.5

2

2.5

3

ND1 ND2 ND3 ND4 ND4l ND5 ND6 Cytb COX1 COX2 ATP6 ATP8

mR

NA

(rel

ativ

e ex

pres

sion

)+ resveratrol (wildtype)+ resveratrol (SIRT1 knockout)

Mitochondrially-encoded ETC genes

Baur et al., Nature (2006)

Haigis and Sinclair, Annu Rev Pathol., 2010

LKB1

Mito poison?Lower ATP SIRT1-independent

SIRT1 activating?Higher ATPSIRT1-dependent

0

5

10

15

20

25

30

35

40

45

50

Standard Diet HFD HFD + Low Resveratrol

HFD + High Resveratrol

ATP

(pmol/m

g protein)

Resveratrol increases ATP in a SIRT1-dependent manner in vivo

Gastrocnemius

p –AMPK (Thr 172)

Total AMPK

SIRT1 KOWT WT SIRT

1 KOWT

SIRT1

KO

WTSIRT1 KO

Standard diet

HFD HFD + High RSV

HFD + Low RSV

0

0.5

1

1.5

2

2.5

Standard diet

HFD HFD + Low RSV

HFD + High RSV

p-A

MP

K/A

MP

K

WTSIRT1 KO

Is AMPK activation SIRT1-dependent?

1st screen of ~18,000 molecules: found 18(best in class - resveratrol)

Discovery and Development of SIRT1 activators (STACs)

Screened 1,000,000 molecules: 6 novel chemical scaffoldsSynthesis of 4000+ STACs

Potency increase 10,000-foldEfficacy in rodent models of type II diabetes and neuroD

2003-4

2005-9

Human safety and efficacy studies with synthetic STACsHuman data to be publicly released June, 2011 (ADA meeting)

2010-11

Milne et al., Nature (2007)

SIRT1

Ac

SIRT1 SIRT1

W

SIRT1

Amino methyl coumarin (AMC) Tryptophan (W)

Ac

Dai et al., JBC (2010)

Supported by an NIH/NIA-Sirtris CRADA to Rafa deCabo

SRT1720

Joseph Baur, Rafa deCabo, Robin Minor

Muscle Disorders

SarcopeniaDisuse Atrophy

Ophthalmic Diseases

AMDDry eye

Others

Inflammatory BowelSepsis

Metabolic Disease

Type 2 DiabetesOthers

Respiratory

COPD

SRT2104 and SRT2979

A. Camins et al., Biochimica et Biophysica Acta (2010)

Dr. Rafa de CaboAlejandro Martín-Montalvo(NIA/NIH)

Joe BaurBeamon Agarwal(U. Pennsylvania)

Yana CenAnthony Sauve(Cornell University, NY)

Sinclair Lab:Ana GomesAngela HafnerAlvin LingBasil HubbardBrian NorthJun LiLuis RajmanNathan PriceSean Armour

Carlos PalmieraAnabela Rolo(U. Coimbra, Portugal)

Guilio Pasinetti(Mt. Sinai, NY)

Marc DonathMarianne Böni-Schnetzler(University Hospital Basil)

Jim EllisVipin Suri and team(Sirtris)

Funding:  Thanks to NIA/NIH, The Glenn Foundation for Medical Research, The Ellison Medical Foundation.