Nir Barzilai M.D. Director: Institute for Aging Research

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• Nir Barzilai M.D.Director Institute for Aging
  Research
• Departments of Medicine
• and Molecular Genetics

Cambridge, September 2003
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Thank you longevity!!!
Longevity-biology of caloric restriction 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.
Longevity-genes Gil Atzmon Ph.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
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.

• 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)

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65 millions year
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Antagonistic Pleiotropy
65 millions year
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The metabolic syndrome of aging
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How do nutrients induce biological effects?
Nutritional (transcription) regulation of
peptides
Leptin
Glucose-insulin/FFA/AA
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How is leptin expression regulated? Hexosamine
Biosynthetic Pathway
--3-6
Glycosylation Sp1 NFkB PGC1
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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

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Is the HPB activated with aging?
Although old CR animals are metabolically similar
to young rats they have increased levels of
muscle hexosamines (plt0.01)
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(No Transcript)
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Does the HBP effect the muscle function of old
CR rats?
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Excess nutrients can induce peripheral insulin
resistance through the HBP!(and more so in
aging)
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Does the HBP affect adipose tissue function of
old CR rats?
PAI-1 expression (Real-time PCR)
Variety of (harmful?) fat-derived peptides are
over expressed through the HBP.
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Is the transcription of PAI-1 by the HBP
associated with increase in plasma levels?
PAI-1 activity U/ml



Glc
Young
Old CR
Artheriosclerosis. 160 117. 2002.
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Is increased availability of FFA effect PAI-1
transcription in humans adipose tissue?
Similar effects have been induced by
hyperglycemia, hyperinsulinemia, and increased
levels of amino acids in humans and rodents
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Which organ is most important toexert the
biological effects of nutrient excess on
longevity?
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(No Transcript)
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Does VF account for the effects of caloric
restriction on insulin action in aging rats?
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Is the removal of VF sufficient to restore
insulin action?
Young SC- VF- CR
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20
10
0
Rd (mg/kg LBM/min)
Diabetes 512951,2002
Plt0.01
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Does knock out of VF prevents diabetes in
Zucker diabetic rats?
DM
80
40
Diabetes 512951,2002
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Nutrients, nutrient sensing, and induction of
fat-derived peptides
PAI-1
50
Gene expression (adjusted by GAPDH)
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40
35
30
25
16
14
12
10
8
6
4
2
0
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How do nutrients induce biological effects?
PAI-1 AGT
IL-6 IL-1 Resistin SAA
Clinical trials are standardizes by fasting
levels, and are Under estimating daily
transcription of peptides!
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You always understand the results of your
experiment!

• But
• You never know the whole truth!

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100
98
92
95
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Lipoprotein particle sizes with age
Heritabilty of lipoprotein size 0.4-0.7!
9.8
9.6
9.4
HDL Particle Size (nm)
LDL Particle Size (nm)
9.2
9.0
8.8
8.6
20.4
60
65
70
75
80
85
90
95
100lt
60
65
70
75
80
85
90
95
100lt
Age
Age
JAMA (October 03 )
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Is lipoprotein sizes associated with HTN or
CVD?(in offspring of centenarians)
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Cholestryl Ester Transfer Protein (CETP)
TESSSESIQSFLQSMIT
Exon 14
ATC
Isoleucine
Valine
GTC
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CETP I 405 V frequency in families with
exceptional longevity
70
60
50
40
Percentage in population
30
20
10
0
II
IV
VV
CETP VV genotype attributes to 18 of longevity
JAMA (October 03 )
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IS CETP genotype associated with its plasma
levels?
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Is lipoprotein sizes a risk for MS ?(in
offspring and spouse of offspring)
Healthy (n221)
60
70
MS (n47)
65
55
60
50
55
45
50
45
40
40
35
35
30
30
large LDL size
large HDL size
JAMA (2003, 290152030-40 )
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APOC3 C(-641)A frequency in families with
exceptional longevity
Percentage in population
CC
CA
AA
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Rate limiting steps for humans longevity
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Barzilai lab
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Revisiting the role of fat mass in the life
extension induced by caloric restriction

• J. Gerontol Barzilai Gupta 1999, 54B89

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
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Rueben Andres Fault!
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Nutrients, nutrient sensing, and induction of
fat-derived peptides
PAI-1
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Gene expression (adjusted by GAPDH)
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_at_
40
35
30
_at_
25

16
14
12
10

8
6
4
2
0
SC
VF
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Probability of survival in caloric restricted
animals
100
Decrease in all causes of deaths
Survival
Months
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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
  longevity-associated 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
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Lipoprotein sizes and Cognitive function (by
MMSE) in centenarians
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Lipoprotein sizes and I405V genotype


Plt0.001 vs. I/I
JAMA (In Press )
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Does leptin resistance effect insulin action
with aging?
Leptin resistance may lead to alteration in body
fat-distribution and insulin resistance!
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Mitochondrial Overproduction of Superoxide
Activates Four Major Pathways of Hyperglycemic
Damage By Inhibiting GAPDH


NADPH
NADH
NADP
NAD
Sorbitol
Fructose
Polyol pathway
Glucose-6-P
GFAT
Glucosamine-6-P
UDP-GlcNAc
Gln
Glu
Hexosamine pathway

NADH
NAD
a
-Glycerol-P
DAG
PKC
DHAP
Protein kinase C pathway

NAD
Methylglyoxal
AGEs
AGE pathway
NADH
1,3-Diphosphoglycerate
Brownlee M. Nature. 2001414813-820.
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Does the HBP affect adipose tissue function of
old CR rats?
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Does leptin fail to regulate VF with aging?
Barzilai JCI 1997, Rossetti Barzilai Nature
1999 Gabriely Barzilai Diabetes 2002, Xma
Barzilai J Gerontol 2002.
Leptin resistance may lead to alteration in body
fat-distribution and to insulin resistance!
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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 34622
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Atzmon et al Horm Metab Res. 2002 34622