I reveived a message indicating that you might not have received this
announcement properly. Thank you
Cambridge Healthtech Institute
Frontiers in Aging Research and Age-Related Diseases
October 5-7, 1998 San Diego, California
Cosponsored by HealthSpan Sciences, Inc.
Chairperson
Dr. Bryant Villeponteau
HealthSpan Sciences, Inc.
Co-Chairpersons
Dr. James Curtsinger
University of Minnesota
Dr. Bruce H. Howard
National Institute on Child Health and Human Development, NIH
Dr. Mark A. Lane
National Institute on Aging, NIH
Speakers
Dr. Julie K. Andersen
University of Southern California
Dr. James R. Burke
Duke University Medical Center
Dr. Allan Butterfield
University of Kentucky
Dr. James Curtsinger
University of Minnesota
Dr. Susan R. Doctrow
Eukarion, Inc.
Dr. John J. Egan
Alteon, Inc.
Dr. Walter Funk
Geron Corporation
Dr. David Harrison
The Jackson Laboratory
Dr. Bruce H. Howard
National Institute on Child Health and Human Development, NIH
Dr. Ting-Ting Huang
University of California, San Francisco
Dr. Thomas E. Johnson
University of Colorado at Boulder and GenoPlex, Inc.
Dr. Mark A. Lane
National Institute on Aging, NIH
Dr. Gordon J. Lithgow
University of Manchester and Council Science of Ageing Initiative (UK)
Dr. Ronald Pero
OXiGENE Inc.
Dr. Henry Rodriguez
National Institute of Standards and Technology
Dr. Michael R. Rose
University of California, Irvine
Dr. Hidetoshi Tahara
National Institute of Environmental Health Sciences and Hiroshima
University School of Medicine (Japan)
Dr. Nikolaos Tezapsidis
Mount Sinai School of Medicine
Dr. Bryant Villeponteau
HealthSpan Sciences, Inc.
Dr. Samuel Ward
University of Arizona and HealthSpan, Inc.
Dr. Woodring E. Wright
University of Texas Southwestern Medical School
=20
Mechanisms of Aging and Therapeutic Targets
Genetic Analysis of Extended Life Span
Molecular Genetics of Aging and Stress Response
Using Selection to Analyze Postponed Aging
Single Genes Retard Mammalian Aging
Defining Replicative Senescence with Array-Based Expression Analyses
Life Extension as Resistance to Stress
Cell Senescence and Telomerase
Regulation of Higher Order Chromatin Structure
Telomerase and Senescence-Associated Genes
Extension of Cellular Life Span
DNA Repair, Cell Senescence, and Aging
Mapping Oxidative Genomic DNA Damage
Oxidative Stress, Aging, and Disease
The Role of Oxidative Stress in Aging and Disease
Salen Manganese Complexes for Age-Associated Disorders
Relevance of Free Radicals and Oxidative Stress to Alzheimer=92s Disease
Genetic Modifiers for Neonatal Lethality and Cardiomyopathies
C. elegans and Oxidative Stress Factor Research
Therapeutic Strategies: General and Age-Related Diseases
Protein-Protein Interactions in Aging and Neurodegenerative Disease
Exploring the Role of Apoptosis in Neurodegeneration
Implications of Presenilins and Microtubules in Alzheimer=92s Disease
Breaking A.G.E. Cross-links
Potential Metabolic Targets
=20
Monday, October 5
9:00-11:00 am Early Registration, Poster and Exhibit Set-up
1:00 Chairperson=92s Welcome and Opening Remarks
Dr. Bryant Villeponteau, President, HealthSpan Sciences, Inc.
Mechanisms of Aging and Therapeutic Targets
1:10 Co-Chairperson=92s Remarks
Dr. James Curtsinger, Professor, Department of Ecology, Evolution and
Behavior, University of Minnesota
1:15 Using Selection to Analyze Postponed Aging
Dr. Michael R. Rose, Professor, Department of Ecology and Evolutionary
Biology, School of Biological Sciences, University of California, Irvine
A genomics tool that has not been sufficiently appreciated is the use of
selection to identify genes and physiological pathways that determine
function. We have used this approach extensively to identify and
analyze the physiological pathways and candidate genes controlling aging
in the fruit fly, Drosophila melanogaster. These methods can be used
with other organisms as well, including outbred mice.
1:50 The Molecular Genetics of Aging and Stress Response
Dr. Gordon J. Lithgow, Lecturer, School of Biological Sciences,
University of Manchester, and Coordinator of the Biotechnology and
Biosciences Research Council Science of Ageing Initiative
The life-span prolongation mutations (age mutations) of Caenorhabditis
elegans are an important system for understanding normal aging and the
physiological consequences of extended life span. An insulin signaling
pathway controls both C. elegans life span and intrinsic stress
resistance. We have demonstrated genetically and pharmacologically that
this pathway controls the expression of molecular chaperone genes,
suggesting that chaperones, in turn, confer both these phenotypes. We
have used the relationship between stress resistance and extended life
span to rapidly screen for novel mutations that confer stress resistance
and indeed have found single-gene mutations that extend life span. In
collaboration with Stuart Kim (Stanford) we are assessing
gene-regulatory alterations in these novel mutant strains using
microarray technology. We are also looking at detrimental side effects
of life-span extension by testing the relative fitness of strains
carrying age alleles and wild-type strains in laboratory-based natural
selection experiments and find that under specific environmental
conditions there is a fitness cost associated with longevity.
2:25 Single Genes That Appear to Retard Mammalian Aging
Dr. David Harrison, Senior Staff Scientist, The Jackson Laboratory
Genes that differ in natural populations were studied, as these may
confer a selective advantage and illustrate how to retard aging rates.=20
Two different loci each increased maximum life spans more than 100 days
(a third the effect of food restriction) with significances P =3D 2.4 and
4 x 10-5 by chance, meaningful even considering multiple testing. Mouse
populations were derived from crosses of four inbred strains. High
heritabilities of life spans, 46 and 51%, occurred only in populations
including unconventional wild mouse strains, MOLD or CAST. Alleles that
retarded aging substantially were not found in the six conventional
strains but only in the two genetically diverse strains recently derived
from wild mice.
3:00 Poster and Exhibit Viewing, Refreshment Break
3:30 Life Extension as Resistance to Stress: Toward the Molecular
Description
Dr. Thomas E. Johnson, Institute for Behavioral Genetics, University of
Colorado at Boulder, and GenoPlex, Inc.
The stress response hypothesis suggests that "interventions that
increase the response to stress offer the potential for effective life
prolongation and increased health." The hypothesis is supported by
extensive data from gerontogene mutants and overexpressing strains in
the nematode Caenorhabditis elegans. Both these data, as well as data
showing that dietary restriction and environmentally induced life
extension are mediated by common processes, will be reviewed. GenoPlex
is applying this methodology to the identification of evolutionarily
conserved genes and processes leading to reduced aging and increased
health span.
4:05 Defining Replicative Senescence with Array-Based Expression
Analyses
Dr. Walter Funk, Staff Scientist, Geron Corporation
Replicative senescence represents the final stage of normal human
cultured cells and has been suggested to model aging processes in vivo.=20
Much of the work on culture senescence has focused on the
characterization of differences in function and expression seen when
comparing young quiescent cultures to replicative senescence. Two
recent avenues of investigation have included the ability of a variety
of insults, such as oxidative stress and oncogene activation, to induce
senescence-like states and the ability of telomerase activation to
extend the replicative life span of cells. In collaboration with
Synteni Corp., we have developed a high-density custom DNA array that
allows us to monitor the expression of 1,000 independent mRNAs and have
used it to assess the expression profiles of cells at replicative
senescence in multiple cell strains and cell types. Comparisons have
been made to a variety of cell stressors and checkpoint arrests and have
allowed us to tightly define and distinguish the senescent phenotype.=20
In addition, normal cells that express telomerase by means of a
transgene (telomerized cells) have been characterized, and a comparison
of their behavior to control populations will be presented. The use of
telomerized cells in cell and gene therapy applications will be
discussed.
4:40 Genetic Analysis of Extended Life Span in Drosophila: Lessons from
Old Flies
Dr. James Curtsinger
Modern molecular techniques make it possible to genetically dissect
complex polygenic characters such as life span. Using Quantitative
Trait Locus (QTL) analysis, we have been able to identify specific,
small chromosomal regions that carry the genes responsible for doubling
life spans in artificially selected lines of flies. Four QTL peaks
account for most of the selection response, one of which is centered at
the structural genes for Cu/Zn SOD. Fine-scale mapping and tests for
pleiotropic effects of the longevity-enhancing genes are in progress.
5:10 Panel Discussion
5:45-7:00 Reception (sponsored by Cambridge Healthtech Institute)
Tuesday, October 6
8:00 am Poster and Exhibit Viewing, Light Continental Breakfast=20
8:30 Co-Chairperson=92s Remarks
Dr. Bruce H. Howard, Chief, Laboratory of Molecular Growth and
Regulation, National Institute on Child Health and Human Development,
NIH
Cell Senescence and Telomerase
8:35 Regulation of Higher Order Chromatin Structure in Cell Senescence
and Aging
Dr. Bruce H. Howard
Cellular senescence involves the cell cycle-dependent reprogramming of
gene expression, one consequence of which is growth arrest despite
mitogenic stimulation. Consistent with the notion that changes in
higher order chromatin structure accompany such reprogramming, the
histone deacetylase inhibitors trichostatin A and sodium butyrate can
induce commitment of mouse and human fibroblasts to a senescence-like
state. To gain further insight into the genetic pathways involved, we
have established a system in which primary mouse skin fibroblasts are
transduced with retrovirus vectors encoding chromatin-modifying
enzymes. These experiments reveal the p300/CBP-associated histone
acetyltransferase PCAF to have a role in senescence pathways.
9:10 Telomerase and Senescence-Associated Genes: Implication of
Cellular Mortality
Dr. Hidetoshi Tahara, Visiting Fellow, Laboratory of Molecular
Carcinogenesis, National Institute of Environmental Health Sciences, and
Associate Professor, Hiroshima University School of Medicine
Telomere shortening is one of the major alterations that occur during
cellular aging in vitro and is generally believed to be associated with
the upregulation of senescence genes. Telomerase, a ribonucleoprotein
enzyme that adds telomere sequence at the end of each chromosome, is
found in most immortal human cells, including cancer cells, but not in
most normal, mortal cells. Introduction of the telomerase catalytic
subunit, hTERT, into human mortal cells that do not have telomerase
activity resulted in functional telomerase activity and extension of
life span in some human cells. These results suggest that telomere
length may, in part, regulate cellular mortality. In addition,
senescence-associated genes not involved in telomerase regulation may
also provide a critical function in cellular senescence and
immortalization of human cells. These other candidate genes will be
reviewed.
9:40 Extension of Cellular Life Span: Implications for Aging and Cancer
Dr. Woodring E. Wright, Department of Cell Biology and Neuroscience,
University of Texas Southwestern Medical School
Chromosomes are capped by structures called telomeres. DNA polymerase
is unable to replicate the ends of linear DNA molecules, and the
ribonucleoprotein telomerase compensates for this by adding telomeric
repeats to the ends of the chromosome. Telomerase is turned off in most
somatic tissues during development, and in its absence telomeres
shorten. The progressive shortening of telomeres ultimately limits the
number of times normal diploid human cells are able to divide. The
recent ability to prevent telomere shortening by constitutively
expressing the catalytic subunit of telomerase in normal diploid cells
and the demonstration that this greatly extends cellular replicative
life span profoundly affects our approaches for the treatments of
genetic defects, diseases of aging, and cancer.
10:15 Poster and Exhibit Viewing, Refreshment Break
10:45 Cell Senescence, Aging, and DNA Repair
Dr. Ronald Pero, Chief Scientific Officer, OXiGENE Inc.
Summary not available at time of printing.
11:20 Development of LMPCR for Mapping Oxidative Genomic DNA Damage
Dr. Henry Rodriguez (presenting), Biotechnology Division, National
Institute of Standards and Technology, and Dr. Steven A. Akman, Wake
Forest Cancer Center
By developing an oxidative-induced DNA damage mapping version of the
LMPCR technique, we investigated in vivo and in vitro frequencies of DNA
base modifications caused by ROS in the human p53 and PGK1 gene. Human
male fibroblasts were exposed to 50 mM H2O2, or purified genomic DNA was
treated with 5 mM H2O2, 100 =B5M Ascorbate, and 50 =B5M, 100 =B5M, or 100=
=B5M
of Cu(II), Fe(III), or Cr(VI) respectively. Damage patterns generated
in vivo were nearly identical to the in vitro Cu(II)-, Fe(III)-, or
Cr(VI)-mediated damage patterns. Cr(VI) treatment damaged several
unique thymine positions. Also, when cells undergo severe oxidative
stress, extra nuclear sites are a major contributor of metals that can
enter the nucleus and enhance DNA damage. The local probability of
H2O2-mediated DNA damage is determined by DNA sequence, chromatin
structure having limited effect. The data suggest a model in which
DNA-metal binding domains accommodate different metal ions. LMPCR is of
significant interest since elucidating the role oxidatively induced DNA
damage has in carcinogenesis can enable one to develop rational
therapeutic interventions.
11:55 Panel Discussion
12:30 Luncheon (sponsored by Cambridge Healthtech Institute)
2:00 Chairperson=92s Remarks
Dr. Bryant Villeponteau
Oxidative Stress, Aging, and Disease
2:05 The Role of Oxidative Stress in Aging and Disease
Dr. Bryant Villeponteau
The aging process affects the progression of many age-related
degenerative diseases. Effective treatments to slow aging may work on
multiple age-related diseases and thus serve to maximize human health
span. Many studies have indicated that oxidative stress plays a
significant role in aging and in age-related diseases. A summary of
some of the data on the role of oxidative stress in aging and disease
will be given. HealthSpan has developed a patent-pending series of
nutraceuticals that work synergistically to reduce oxidative stress and
inflammation. The nutraceuticals act through well-known biochemical
pathways and may slow the basic aging rate so as to alter the
progression of multiple age-related diseases.
2:40 Salen Manganese Complexes: Combined SOD/Catalase Mimics with Broad
Pharmacological Efficacy
Dr. Susan R. Doctrow, Vice President, Research, Eukarion, Inc.
Salen manganese complexes are multifunctional catalytic scavengers of
reactive oxygen species (ROS), exhibiting both superoxide dismutase and
catalase activities. They are effective in vivo in models for e.g.,
Parkinson=92s disease, stroke, ARDS, multiple sclerosis, and myocardial
infarction. Overall, this is a class of compounds with a novel mode of
action and potential therapeutic utility in the many diseases in which
ROS have been implicated, including age-associated disorders.
3:15 Poster and Exhibit Viewing, Refreshment Break
3:45 Free Radicals and Oxidative Stress: Relevance to Alzheimer=92s
Disease
Dr. Allan Butterfield, Professor, Department of Chemistry, Center of
Membrane Sciences, and Sanders-Brown Center on Aging, University of
Kentucky
Substantial evidence exists for oxidative stress in Alzheimer=92s disease
(AD) brain. Although several sources of oxidative stress may be
operative in AD brain, the role of amyloid b-peptide (Ab), the chief
constituent of senile plaques in AD brain, and its membrane-based
reaction products in free radical-induced oxidative stress has been the
subject of intense investigation in our laboratory. This talk will
summarize some of these Ab-induced markers of oxidative stress and
include results from antioxidant studies that show modulation of the
effects of Ab on brain cell oxidative stress and neurotoxicity.
4:20 Genetic Modifiers for Neonatal Lethality and Cardiomyopathies in Mn
SOD Mutant Mice
Dr. Ting-Ting Huang, University of California, San Francisco
The role of oxidative damage in the aging process and in a number of
age-related degenerative diseases has been well documented. Mutant mice
(Sod2-/-) lacking the mitochondrial superoxide metabolizing enzyme, Mn
superoxide dismutase (MnSOD), represent an animal model with increased
mitochondrial superoxide radicals, accelerated tissue damage, and early
demise. When the mutant mice are placed on an outbred (CD1) genetic
background, they have a mean life span of 5.4 days and develop a severe
metabolic acidosis (ketosis) at birth and dilated cardiomyopathy by 4
days of age. Reduced [Fe-S] cluster containing enzymes were also
observed in different tissues of the mutant mice. When the mutant mice
are placed on a C57BL/6J background, a more severe phenotype is
observed. Half of the knockout mice die in utero starting at gestation
day 15, and the live-born knockout mice have a mean life span of 1.5
days. In addition, cardiomyopathy is observed at birth. However, when
the mutant mice are generated as B6D2 (DBA/2J) F2 or (B6D2 F1) B6
backcross animals, they survive for up to 21 days and the long-lived
(>15 days) knockout mice have a mild metabolic acidosis and almost no
signs of cardiomyopathy. The data indicate that genetic components that
cosegregate with the long-lived population in the F2 and backcross Sod2
knockout mice have the ability to decelerate tissue damage and
consequently, prolong the life span.
4:55 Metabolic Changes in Long-Lived Mutants of the Nematode C. elegans
Dr. Samuel Ward (presenting), University of Arizona and HealthSpan,
Inc., and Dr. Wayne Van Voorhies, University of Arizona
By directly measuring CO2 production from worms growing under optimal
conditions, we have found that metabolic rate is reduced in all of the
long-lived mutants that we have tested. When a suppressor mutation that
returns the life span to normal is introduced into one of these strains,
the metabolism is also restored to normal. These results are consistent
with the known biochemical defects in these genes and suggest that
reduced oxidative metabolism is the cause of the extended life span.=20
The most likely biochemical mechanism is that this reduces oxidative
stress in the worms.
5:30 Panel Discussion
Wednesday, October 7
8:00 am Poster and Exhibit Viewing, Light Continental Breakfast
8:30 Co-Chairperson=92s Remarks
Dr. Mark A. Lane, National Institute on Aging, Intramural Research
Program, Gerontology Research Center
Therapeutic Strategies: General and Age-Related Diseases
8:35 Protein-Protein Interactions in Aging and Neurodegenerative
Disease: Polyglutamine Domain Proteins as a Model System
Dr. James R. Burke, Assistant Professor, Medicine and Neurology, Deane
Laboratory, Duke University Medical Center
Abnormal protein-protein interactions leading to aggregation are a
hallmark of aging and neurodegenerative diseases including Alzheimer=92s,
Parkinson=92s, and Huntington=92s diseases. Expanded polyglutamine domai=
n
proteins are an excellent model system because (1) they cause eight
neurodegenerative diseases, (2) age-of-onset and disease expression are
determined by the length of the polyglutamine domain, (3) expression of
polyglutamine domain proteins in cultured cells and transgenic animals
demonstrates polyglutamine length-dependent aggregation and cell death
mimicking the effects in humans, and (4) cell death is independent of
the protein sequence surrounding the polyglutamine domain. We have used
polyglutamine-green fluorescent protein fusion proteins to track the
intracellular fate of polyglutamine domain proteins and to characterize
interacting proteins.
9:10 A Model System for Exploring the Role of Apoptosis in
Neurodegeneration
Dr. Julie K. Andersen, Assistant Professor, Andrus Gerontology Center,
University of Southern California
The baculoviral protein p35 is a general irreversible inhibitor of
caspase enzymes that appear to be major molecular mediators of
apoptosis. Transgenic lines have been generated in which p35 is
expressed neuronally. One particular line has elevated levels of p35
expression in pyramidal neurons throughout the hippocampus and in the
granular layer of the cerebellum. Cerebellar granular cells isolated
from these are more resistant to apoptosis induced by various
mechanisms. We are using these lines to explore the possible role of
caspases in apoptosis associated with beta-amyloid induced cell death in
hippocampal neurons as a model for Alzheimer=92s disease.
9:45 Presenilins and Microtubules: Implications in Alzheimer=92s Disease
Dr. Nikolaos Tezapsidis, Assistant Professor of Research in Psychiatry,
Mount Sinai School of Medicine
In an attempt to identify a function for the presenilins (PS-1 and
PS-2), a search is under way for proteins able to bind them and
consequently mediate their action. The strategy involves the screening
of a brain expression library with probes comprising heterologous
regions of PS-1 and PS-2. Candidates are characterized for their
binding properties to PS-1 and PS-2 using in vitro assays, and the
perturbations of these interactions by mutations linked to familial
Alzheimer=92s disease (AD) are evaluated. Specific interactors are
further analyzed for their efficacy to alter the Ab peptide levels in
cell culture experimental models. One of the binding proteins was
identified as a microtubule-interacting protein. A potential role of
the presenilins in cytoskeletal architecture and dynamics will be
discussed along with the evaluation of novel therapeutic targets for AD.
10:15 Poster and Exhibit Viewing, Refreshment Break
10:45 Breaking A.G.E. Cross-links: Reversing Complications of Aging
Dr. John J. Egan, Senior Director, Preclinical Research, Alteon, Inc.
Glucose reacts with proteins through nonenzymatic glycosylation to form
sugar adducts, "A.G.E.s" (advanced glycation endproducts), containing
a-dicarbonyl motifs. These extremely reactive motifs form covalent
cross-links with amines on adjacent proteins. Cross-linking, which is
accelerated in the diabetic state, is postulated as central to the
development and the severity of complications associated with diabetes
and aging. In diabetic rats, ZS hypertensive rats, aging dogs, and
aging monkeys, lasting improvements are observed in reversing
complications of the cardiovascular system following short-term
treatment (weeks) with a compound designed to chemically cleave
carbon-carbon bonds within the a-dicarbonyl motif. Results indicate
that reversal of some complications of aging such as cardiovascular
stiffening can readily be achieved following breakage of A.G.E.
cross-links.
11:20 Potential Metabolic Targets for Interventions to Mimic Effects of
Calorie Restriction
Dr. Mark A. Lane (presenting), Dr. Donald K. Ingram, and Dr. George S.
Roth, Intramural Research Program, Gerontology Research Center, National
Institute on Aging, NIH
Recent evidence from monkey studies suggests that calorie restriction
(CR), which extends life span in rodents, might have beneficial effects
in long-lived primate species, perhaps even humans. A 30% reduction in
calories would not be well tolerated in most people, highlighting the
importance of discovering the "anti-aging" mechanism of CR. Our
laboratory is focusing on glucose and insulin metabolism in the CR
paradigm. We have shown that 2 deoxy-D-glucose, a structural analogue
of glucose, mimics certain biological hallmarks of CR such as reduced
body weight, temperature, and fasting insulin. Another line of work has
shown that genes related to life span in nematodes that share homology
with mammalian insulin signaling genes may be altered during CR.
12:00 Chairperson=92s Closing Remarks
Dr. Bryant Villeponteau
12:05 Close of Conference
