Transcript Successful Longevity

Nir Barzilai M.D.
Director: Institute for Aging Research
Departments of Medicine
and Molecular Genetics
Cambridge, September 2003
Thank you longevity!!!
Longevity-biology of caloric restriction
Longevity-genes
Xiao-Hui Ma M.D
Xiao-man Yang M.D.
Radhika Muzumdar M.D.
Bing-Qian Liu M.D
Ilan Gabriely M.D.
Patricia Vuguin M.D.
Gil Atzmon Ph.D.
Program project
Jill Crandall M.D.
Meredith Hawkins M.D.
Harry Shamoon M.D.
Luciano Rossetti M.D.
Michael Brownlee M.D.
Silvana Obici M.D.
Bill Greiner R.N.
Magda Gabriely M.D.
Debra Davidson M.Sc.
Clyde Schechter M.D.
Richard Lipton M.D.
Ms. Marion Maze
Ms. Linda Radner
Other Collaborators
Alan Shuldiner M.D. (UMD)
Braxton Mitchell Ph. D. (UMD)
Hassy Cohen M.D. (UCLA)
Tom Perls M.D. (BU)
Gad Rennert M.D., Mp.H.(Technion)
Amiela Globerson Ph.D. (Ben Gurion)
Norman Fleischer M.D (Einstein)
Philip Scherer Ph.D. (Einstein)
Becky Simmons M.D. (Penn)
65 millions year
50 years
Antagonistic Pleiotropy
65 millions year
50 years
The metabolic syndrome of aging:
Insulin resistance High insulin
Obesity/abdominal obesity
Type 2 diabetes mellitus
Dyslipidemia
Low HDL, ‘small’ LDL
Hypertension
AGT
Inflammation
IL-1, IL-6, CRP
Endothelial dysfunction
Thrombosis
PAI-1
ASCVD
Cancer
How do nutrients induce biological effects?
Nutritional (transcription) regulation of peptides
Leptin
2 am
Glucose-insulin/FFA/AA
8 am
Barzilai JCI 1997,
Rossetti & Barzilai Nature 1999,
Gabriely & Barzilai Diabetes 2002,
Xma & Barzilai J Gerontol 2002.
Leptin fails to regulate body fat distribution,
insulin action, and endocrine functions with aging
8 am
How is leptin expression regulated?
Hexosamine Biosynthetic Pathway
Glucose
AA (Glutamine)
Glucosamine
Glc-1-P
Glucose-6-P
GFAT
1%
UDP- Glc
--3-6%
GlcN-6-P
F -6-P
FFA
Triose -P
UDPGlcNAc
Glycogen
Glycolysis
Glycosylation
Sp1
NFkB
PGC1
FFA
How can a pathway serve as
‘nutrient sensing’?
• Should serve as a switch “on” or “off”.
• Should be a small pathway that can be amplified by
several folds.
• Should be linked to a biological effect (transcription).
• Should exist in every organ, although may be distinct.
• Should apply to all models of CR
Is the HPB activated with aging?
Muscle UDPGlcNAc/UDPGlc
**
*
3
2
1
0
Young
Old AL
Old CR
Although old CR animals are metabolically similar to young rats
they have increased levels of muscle hexosamines (p<0.01)
Does the HBP effect the muscle
function of old CR rats?
3 mo (n=5) and 21 mo (n=5) CR S-D rats
Insulin-3 mU/kg/min
GlcN-7 µM/kg/min/or Saline
Rd (mg/kg/min)
30
****
25
Young/old CR saline control
****
UDP-GlcNAc 20.8 nmol/g muscle
20
****
Young
Old
15
10
5
P<0.001
UDP-GlcNAc 17.7 nmol/g muscle
290
210
230
250
270
90
110
150
190
30
50
70
0
Time (min.)
Excess nutrients can induce peripheral
insulin resistance through the HBP!
(and more so in aging)
Does the HBP affect adipose tissue
function of old CR rats?
PAI-1 expression
(Real-time PCR)
14
12.5
12
10
8
Young
Old
6
4
3.2
2.2
2
1
0
Saline
GlcN
Variety of (harmful?) fat-derived peptides are over expressed
through the HBP.
Is the transcription of PAI-1 by the HBP
associated with increase in plasma levels?
PAI-1 activity U/ml
30
*
25
20
15
*
*
10
5
0
Glc
+
-
Young
Artheriosclerosis. 160: 117. 2002.
+
-
Old CR
ß-actin
PAI-1
30
(relative to baseline)
Gene expression level
Is increased availability of FFA effect PAI-1
transcription in humans adipose tissue?
20
10
0
Liposyn
-
+
Hawkins et al
Similar effects have been induced by
hyperglycemia, hyperinsulinemia, and increased
levels of amino acids in humans and rodents
Which organ is most important to
exert the biological effects of
nutrient excess on longevity?
Does VF account for the effects of caloric
restriction on insulin action in aging rats?
Young
Old SC-
Old VF-
Old CR
500
400
300
200
100
0
Body Weight (g)
Diabetes; 51:2951, 2002
Is the removal of VF sufficient
to restore insulin action?
Young
SC-
VF-
CR
30
*
20
10
0
Rd (mg/kg LBM/min)
*P<0.01
Diabetes; 51:2951,2002
Does ‘knock out’ of VF prevents diabetes
in Zucker diabetic rats?
%DM
120
~80%
100
80
ZDVFZDVF+
60
40
~40%
20
0
2 mo
3 mo
Diabetes; 51:2951,2002
4 mo
5 mo
6 mo
PAI-1
16
14
12
10
8
6
4
2
0
$$
12
10
8
*
Gene expression (adjusted by
Gene expression (adjusted by
GAPDH)
GAPDH)
Gene expression (adjusted by
GAPDH)
$$@
Saline
Glucose
Insulin
GlcN+In
$$
@
Saline
Glucose
Insulin
GlcN+In
20
$
0
$$
10
0
TNF- 
5
4
3
2
1
0
$$$
30
4
6
Resistin $$$
$
40
6
2
60
50
$$
Angiotensinogen
20
18
16
14
12
10
8
6
4
2
0
@
Saline
Glucose
Insulin
GlcN+In
Gene expression (adjusted by
GAPDH)
$@
Saline
Glucose
Insulin
GlcN+In
Leptin
Saline
Glucose
Insulin
GlcN+In
$@@
$$
$$
Acrp30
Gene expression (adjusted by
GAPDH)
50
45
40
35
30
25
Gene expression (adjusted by GAPDH)
Gene expression (adjusted by
GAPDH)
Nutrients, nutrient sensing, and induction of fat-derived peptides
16
14
12
10
8
6
4
2
0
Saline
Glucose
Insulin
GlcN+In
@
$$
$$
How do nutrients induce biological effects?
FAT+Nutritional (transcription) regulation of peptides
Leptin
2 am
PAI-1
AGT
IL-6
IL-1
Resistin
SAA
8 am
Glucose/insulin/FFA
Clinical trials are standardizes by fasting levels, and are
Under estimating daily transcription of peptides!
8 am
You always understand the results of your
experiment!
But
You never know the whole truth!
100
98
92
95
Lipoprotein particle sizes with age
9.8
21.6
9.6
21.4
LDL Particle Size (nm)
HDL Particle Size (nm)
Heritabilty of lipoprotein size 0.4-0.7!
9.4
9.2
9.0
21.2
21.0
Probands
20.8
8.8
20.6
8.6
20.4
60
65
70
75
80
Age
85
90
95 100<
Control
Offspring
60
65 70 75 80 85
90 95 100<
Age
JAMA (October 03 )
Is lipoprotein sizes associated with HTN or CVD?
(in offspring of centenarians)
60
*
70
*
60
50
Healthy (n=209
HTN (n=64)
CVD (n=20)
50
40
40
30
30
20
20
% large HDL size
*P<0.003
% large LDL size
JAMA (October 03 )
Cholestryl Ester Transfer Protein (CETP)
Exon 14
TESSSESIQSFLQSMIT
Homozygous
II
= Isoleucine
GTC
= Valine
A
A
A
ATC
OR
G
Heterozygous
IV
G
OR
G
Homozygous
VV
CETP I 405 V frequency in families
with exceptional longevity
Control
Offspring
70
Percentage in population
Probands
60
50
40
30
*
20
***
10
0
II
IV
VV
CETP VV genotype attributes to 18% of longevity
JAMA (October 03 )
IS CETP genotype associated with its plasma
levels?
2.4
I/I
I/V
V/V
2.2
2
1.8
*
1.6
1.4
1.2
1
CETP le vels (ug/ml)
*P<0.02
JAMA (October 2003 )
Is lipoprotein sizes a risk for MS ?
(in offspring and spouse of offspring)
70
***
65
60
***
Healthy
MS (n=47)
(n=221)
55
60
50
55
50
45
45
40
40
35
35
30
30
% large HDL size
***P<0.001
% large LDL size
JAMA (2003, 290;15;2030-40 )
APOC3 C(-641)A frequency in families
with exceptional longevity
Control
Offspring
70
Probands
Percentage in population
60
50
40
30
20
***
***
10
0
CC
***P<0.001
CA
AA
Rate limiting steps for humans longevity
100
80
60
Alive
cancer
40
CVD
CR
20
Telomeres
Mitochondria
0
60
80
100
120
140
Age
Barzilai lab
Revisiting the role of fat mass in the life
extension induced by caloric restriction
•
J. Gerontol Barzilai & Gupta; 1999, 54:B89
The role of fat mass as causative of age-related diseases
leading to morbidity and mortality is well accepted.
Type 2 DM is a striking example.
...This is not a novel hypothesis
Mounting evidence supports the notion that fat mass is inert
and plays no role, as compared with nutrients per se in CR.
…This hypothesis should not be revisited
Rueben Andres
Fault!
*
@
$@
$$
$$
SC
Leptin
12
10
8
$$
*
$$
*
@
$$@
$$
0
VF
$$
10
SC
VF
VF
TNF- 
6
5
$$$
0
Acrp30
Saline
Glucose
Insulin
GlcN+In
4
*
3
2
$@@
$$
$$
1
0
SC
$
20
SC
Gene expression (adjusted by
Gene expression (adjusted by
GAPDH)
GAPDH)
Gene expression (adjusted by
GAPDH)
Saline
Glucose
Insulin
GlcN+In
$$$
30
@
$
Angiotensinogen
20
18
16
14
12
10
8
6
4
2
0
60
50
2
VF
Resistin
Saline
Glucose
Insulin
GlcN+In
40
6
4
@
Saline
Glucose
Insulin
GlcN+In
Gene expression (adjusted by
GAPDH)
16
14
12
10
8
6
4
2
0
Saline
Glucose
Insulin
GlcN+In
SC
VF
Gene expression (adjusted by
GAPDH)
PAI-1
50
45
40
35
30
25
Gene expression (adjusted by GAPDH)
Gene expression (adjusted by
GAPDH)
Nutrients, nutrient sensing, and induction of fat-derived peptides
16
14
12
10
8
6
4
2
0
Saline
Glucose
Insulin
GlcN+In
@
$$
$@
$
$
SC
VF
Probability of survival in
caloric restricted animals
100%
Survival
Decrease in all causes
of deaths
Months
Human longevity genes
We hypothesize that to get to age 100 (1/10,000) one needs longevity assurance genes.
Offspring inherit longevity traits from their parents, and these may be a platform for longevityassociated traits.
We have collected over 1000 samples! (At their homes)
Our population of (Ashkenazi) Jews is genetically homogenous, increasing the likelihood of genetic
discoveries.
Funding by AECOM the National Institute of Aging and the Ellison medical Foundation
Lipoprotein sizes and Cognitive function (by MMSE)
in centenarians
100
21.5
80
21.2
60
20.9
40
20.6
20
20.3
0
20
% large LDL size
MMSE>25
MMSE<25
LDL size (nm)
Lipoprotein sizes and I405V genotype
70
60
*
*
60
50
I/I
I/V
V/V
50
40
40
30
30
20
20
% large HDL size
*P<0.001 vs. I/I
% large LDL size
JAMA (In Press )
Does leptin resistance effect
insulin action with aging?
% suppression of hepatic glucose production
Leptin vs. pair-fed
0
-20
*
-40
-60
-80
-100
Young
Gabriely et al Diabetes 51:1016, 2002
Old CR
Leptin resistance may lead to alteration in
body fat-distribution and insulin resistance!
Mitochondrial Overproduction of Superoxide Activates Four
Major Pathways of Hyperglycemic Damage By Inhibiting
GAPDH
NADP +
NADPH
Glucose
NAD +
NADH
Sorbitol
Fructose
Polyol pathway
Glucose-6-P
GFAT
Glucosamine-6-P
Fructose-6-P
Gln
UDP-GlcNAc
Glu
Hexosamine pathway
NADH
DHAP
NAD +
 -Glycerol-P
DAG
PKC
Protein kinase C pathway
Glyceraldehyde-3-P
NAD +
GAPDH
NADH
O2– •
Methylglyoxal
AGEs
AGE pathway
1,3-Diphosphoglycerate
Brownlee M. Nature. 2001;414:813-820.
Does the HBP affect adipose tissue
function of old CR rats?
Leptin expression
(Real-time PCR)
12
11
10
8
Young
Old
6
4
3.2
2.3
2
1
0
Saline
GlcN
Does leptin fail to regulate VF with aging?
% decrease
Leptin vs. pair-fed
0
-10
-20
*
-30
-40
-50
-60
Young
Barzilai JCI 1997,
Rossetti & Barzilai Nature 1999
Gabriely & Barzilai Diabetes 2002,
Xma & Barzilai J Gerontol 2002.
Old CR
Leptin resistance may lead to alteration in
body fat-distribution and to insulin resistance!
Out off approximately 7,000 full-length sequences
and approximately 1,000 EST clusters
2570 were expressed
200 were up/down- regulated in
each chip
Atzmon et al Horm Metab Res. 2002 34:622
Ge ne
Ins ulin-induce d grow th-re s pons prote in (CL-6)
Be ta 3-adre ne rgic re ce ptor
Phos phoe nolpyruvate carboxyk inas e (GTP) PEPCK
PPAR-gam m a
Horm one s e ns itive lipas e
Ins ulin-lik e grow th factor I
Fatty acid trans porte r
Thioe s te ras e II
M itochondrial 3-hydroxy-3-m e thylglutaryl-CoA s ynthas e
Low m ole cular w e ight fatty acid binding prote in
GLUT1 = glucos e trans porte r 1
Lipopolys accharide binding prote in
Lys os om al acid lipas e = intrace llular hydrolas e
Fatty acid s ynthas e
Type II cAM P-de pe nde nt prote in k inas e re gulatory s ubunit
Adipocyte horm one -s e ns itive cyclic AM P phos phodie s te ras e
Pote ntial-s e ns itive polys pe cific organic cation trans porte r
Re tinol-binding prote in (RBP) ge ne , e xon 5
Ste roidoge nic acute re gulatory prote in
Grow th horm one re ce ptor
Chape ronin 60 (Hs p60) and chape ronin 10 (CPN10) ge ne s , nucle ar
ge ne s e ncoding m itochondrial prote ins
Aquaporin 7
Angiote ns inoge n
Glutathione -de pe nde nt de hydroas corbate re ductas e
M HC clas s II antige n RT1.B-1 be ta-chain
Tricarboxylate carrie r
Thyroid s tim ulating horm one re ce ptor
Phos phodie s te ras e I
Wate r channe l aquaporin 3 (AQP3)
Carbonic anhydras e II
Wis tar-Kyoto (He ide lbe rg) angiote ns in conve rting e nzym e
GST
Na-K-Cl cotrans porte r (Nk cc1)
Alpha-2-u globulin
Glutathione S-trans fe ras e Yc1 s ubunit
Wis tar trans form ing grow th factor be ta-3
Live r glutathione S-trans fe ras e Yc s ubunit
Polym e ric im m unoglobulin re ce ptor
Alk aline phos phatas e
M ultiple incre as e
Pathw ays
In VF
In SC
5.9
5
4.3
4.1
3.5
Glucos e
3.2
hom e os tas is
3.2
Ins ulin action
156
and lipid
21.1
m e tabolis m
10.1
9.8
9.7
4.6
3.9
7.4
5.7
5.6
5.2
5
5
4.8
-
4.3
4.5
4
3.6
3.6
3.6
3.5
-
20.8
11.8
7.4
6.1
5.7
5.3
4.7
4.7
4.4
4
3.9
Ce llular
m e tabolis m and
othe r
Atzmon et al
Horm Metab Res. 2002 34:622