NIH Meeting on Rat
Model Priorities
May 3, 1999
TABLE OF CONTENTS
Foreword Executive Summary
Report Agenda Rosters
FOREWORD
The National Institutes of Health (NIH), realizing the
potential of rat models in understanding basic biology and human health and
disease, launched the Rat Genome Program in 1995, followed by the Rat EST
Program in 1997. These two programs, which are coordinated by the National
Heart, Lung, and Blood Institute (NHLBI), are funded by 13 Institutes and
Centers at NIH, and have produced a wide variety of resources. In addition,
these programs have begun to provide a powerful tool to link and capitalize
upon the data and resources of other model organisms and the human.
We live in an era of extraordinary opportunities and
unprecedented scientific discovery. New developments in genomics, genetics,
drug discovery, stem cell research, bioengineering, and other fields are
creating opportunities for revolutionary changes in the practice of medicine.
The purpose of the Rat Model Priority Meeting was to discuss, within this
context, the opportunities that rat models offer and the investments that are
needed to capitalize on these opportunities. The major issues that were
addressed were: where does rat fit in the broader scientific picture, what
unique value does the rat model provide, what are the key areas of opportunity
for investment, and what will be the impact of these proposed investments.
Participants were charged by Dr. Harold Varmus, Director of NIH, to prepare a
report that contains a summary of the major themes and recommendations, a sense
of priorities, and a practical look at costs.
The workshop was structured to enable as much work as
possible to be done in advance of the meeting (including the use of a Web site
that provided a forum for posting predefined questions and the answers provided
by the scientific community) and was designed for maximal interaction. The
report from this meeting, held May 3, 1999 on the campus of NIH and involving
over forty distinguished scientists, can be found on the NHLBI Web site at
http://www.nhlbi.nih.gov/meetings/index.htm.
EXECUTIVE
SUMMARY
I. BACKGROUND
The rat model provides important strengths for the
study of human health and disease. The large number of inbred rat models and
the vast amount of data (physiological, behavioral, biochemical, cellular,
pharmacological, and toxicological, etc.) provide a superb platform on which to
build the genetic and genomic tools and resources to delineate the connections
between genes and biology. Importantly, in many instances, the rat is the most
appropriate experimental model of human disease.
The National Institutes of Health (NIH), realizing the
potential of rat models in understanding basic biology and human health and
disease, launched the Rat Genome Program in 1995, followed by the Rat EST
Program in 1997. These two programs, funded by 13 Institutes and Centers at
NIH, have produced a variety of basic genomic resources. In addition, these
programs have begun to provide a powerful tool to link to and capitalize upon
the data and resources for both other model systems and the human.
An era of extraordinary opportunities and
unprecedented scientific discovery now presents itself. New developments in
genomics, genetics, drug discovery, stem cell research, bioengineering, and
other fields are creating opportunities for revolutionary changes in the
practice of medicine. The rat model must be poised to take advantage of the
opportunities available, with suitable investments to capitalize on these
opportunities.
As a consequence, the NIH Director, Dr. Harold Varmus,
asked the National Heart, Lung, and Blood Institute (NHLBI) to convene a
meeting to discuss the opportunities and prioritize the needs to fully take
advantage of rat models. On May 3, 1999 a meeting was held on the campus at
NIH.
II. NEEDS AND OPPORTUNITIES
Several needs and opportunities were identified, both
through the responses to questions posted on the Rat Community Forum
(http://goliath.ifrc.mcw.edu/RCF, this Web page is no longer available)
and at the meeting by the participants. These needs and opportunities were
grouped into the two areas of biology/physiology and genomic
infrastructure.
A. BIOLOGY/PHYSIOLOGY:
1. Germ-line Modification
The need for access to the germ-line of the rat in
order to produce informative mutants is critical. The availability of gain of
function (over-expression; knock-in) and loss of function (knock-out) mutations
in the rat will be necessary for the functional characterization of genes.
Correct assignment of quantitative trait loci (QTL) to genes will require
verification from loss of function mutations. Although the production of
transgenic rats is now routine, the creation of loss of function mutations
(knock-out) or gene replacement (knock-in) by homologous recombination in
embryonic stem (ES) cells has not yet been possible in the rat. The gene
replacement strategy is essential, as it provides the means to ultimately study
the "natural" mutations that exist within the various rat models of common
diseases.
2. National Rat Genetic Resource Center
The genetic integrity of, and access to, important rat
strains is maintained by an international effort that depends on the personal
good will of many individuals. This effort is inherently inefficient and
susceptible to the vicissitudes of funding and local interests. A much more
robust approach to the problem of genetic integrity, microbiological quality
and distribution of these valuable models would be the National Rat Genetic
Resource Center, which was proposed in August, 1998 at the NIH Rat Model
Repository Workshop.
3. Rat Genome Database
NIH has issued a request for applications (RFA) for a
Rat Genome Database (RGD) to establish a database that will collect,
consolidate, and integrate data generated from ongoing rat genetic, genomic,
and related research efforts, and to make these data widely available to the
scientific community. The applications were due on April 30, 1999 and an award
is expected September 30, 1999. The participants strongly endorsed this plan
for NIH to implement an RGD.
4. Interaction with the NIH Mouse Mutagenesis and
Phenotyping Program
Mutagenesis and phenotyping of rat models, in parallel
with the NIH mouse mutagenesis and phenotyping program, will allow significant
interaction and collaboration on developing new rodent models, with comparable
characterization, of human disease. Random mutagenesis can offer the rat model
an alternative to gene targeting strategies, as many of the induced mutations
are loss of function. In addition, if rat and mouse mutagenesis and phenotyping
programs are established together, each can benefit from the others experience
-- the mouse model researchers in biological phenotyping and the rat model
researcher in genetic manipulation, as well as the opportunity to compare and
contrast the biology between these two model organisms.
5. Strengthening the Rat Model User Research
Community
A major success of the meeting was catalyzing the
formation of an interactive research community or rat-users. Recommendations to
strengthen this research community included the continued use of the Rat
Community Forum web site, a series of stand-alone rat genomic/genetic-based
meetings, and a number of rat genomic/genetic meetings in conjunction with
mouse and human genetics meetings.
B. GENOMIC INFRASTRUCTURE:
1. Rat EST Project (REST)
The REST project has generated more than 93,000 ESTs
derived from 12 different normalized cDNA libraries. These normalized libraries
combined with a serial subtraction strategy have provided an unprecedented
level of efficiency with respect to identifying unique rat genes, a set of
27,000 Unigene clusters. This is a marked improvement over the 15,000 mouse
Unigene clusters generated from more than 300,000
ESTs. However, workshop participants expressed the view that more comprehensive
coverage (90%) would offer significant advantages and opportunities by
facilitating gene discovery and providing sequence links to the human and mouse
genomic sequence. These sequence links will assist the human and mouse
communities in assigning gene function by providing connections to the wealth
of physiological data in the rat.
2. SNP Discovery and Mapping
The density of markers currently available is nearly
an order of magnitude short of the number of markers that will be required to
positionally clone genes from the mapped positions. To overcome this
limitation, the participants suggested constructing a 3rd generation
genetic map using single nucleotide polymorphisms (SNPs) at a resolution of 1
SNP per 100 kilobases. These markers will also facilitate construction of
physical map (rough-draft and sequence-ready).
3. BAC Clone Resource
Physical mapping reagents exist that are useful for
initiating individual positional cloning projects and are being used by many
laboratories around the world. However, with the change in sequencing capacity
in both the public and the private sector it is important to consider if these
reagents position the rat for the sequencing of its full genome, once the mouse
genome is sequenced. Therefore, there is a need to develop a BAC library with
15-fold coverage made from multiple sources of DNA cut using a variety of
restriction enzymes or random shear techniques while still maintaining an
average insert size greater than 150 kilobases. The currently available BAC/PAC
resources have an average insert size less than the optimal 150 kilobases.
4. Genomic Sequence
Within less than two years the sequencing capacity of
the publicly funded US and international genomics communities are likely to
pass that needed to sequence a mammalian genome to 10-fold redundancy in one
year. The rat should be positioned for genomic sequencing immediately after, or
in parallel, with the mouse.
III. RECOMMENDATIONS/ PRIORITIES
After a thorough discussion of the needs and
opportunities, workshop participants made four major recommendations, which are
listed in priority order:
1. Germ-line Modifications ($8.5 million
over 5 years)
Germ-line modification in the rat is critically
important to assigning gene function to specific genes and to identifying gene
alterations responsible for specific phenotypes. However, germ-line
modification in the rat is limited. The highest priority should be to overcome
these limitations, thorough the use of the following strategies: (a) The
development of nuclear transfer in the rat is an important priority that needs
to be met ($5 million over 5 years). (b) The development of dominant negative
mutations by pronuclear injection should be encouraged ($500,000 per year for 5
years). (c) Cryopreservation of zygotes and sperm will help to alleviate the
storage and transportation problems ($750,000 over 5 years). (d) The generation
of in vitro fertilization techniques in the rat, including intracytoplasmic
sperm injection (ICSI), will help the use of cryopreserved sperm by allowing
suboptimal sperm to lead to viable offspring ($250,000 over 2 years).
2. Additional Genomic Resources ($30
million over 3 years)
a. EST Development ($2.5 million per year for 3
years)
Participants recommended that the goal of the Rat
EST Project be expanded to achieve 90% coverage of all rat genes. Broader
coverage will greatly accelerate efforts to identify genes and elucidate their
functions in advance of having the complete genomic sequence.
b. SNP Discovery ($6.5 million over 3 years)
Developing a SNP map of the rat is a high priority,
as it is required to accelerate the identification of genes (through positional
cloning) responsible for complex, common diseases, as well as facilitate the
construction of a physical map.
c. BAC Clone Resources ($1 million for 1 year)
Sequencing the rat genome should be placed at very
high priority as the third mammalian genome to be sequenced after man and
mouse. The key prerequisite for such an effort is a high quality well
characterized, and deeply redundant BAC library. This library should have large
inserts (average size greater than 150 kilobases), be redundant to a depth of
15-fold or greater, and ideally be produced by more than one restriction enzyme
or physical shearing.
d. Pilot Sequencing ($15 million over 3 years)
A complete sequence of the rat genome will provide
the critical substrate for understanding the molecular basis of biological
function and pathology. A coordinated and systematic approach will be most cost
effective. It is however not clear how to optimally sequence the rat given the
existence of the sequence for mouse and man, a rich array Unigene models from
all three species, and the need to address further genomes. It may be that some
form of draft sequence in the rat would strike the best cost/benefit balance.
To address this question, a small number of rat genome sequencing pilot
studies, each covering a few megabases, should be performed.
3. National Rat Genetic Resource Center
($35 million over 5 years)
Establish this critical resource to maintain and
distribute standard rat models as recommended in the NIH Rat Model Repository
Workshop report (http://www.nhlbi.nih.gov/nhlbi/
sciinf/model/ratmodel.htm) and also explore creative mechanisms for its
maintenance beyond the initial five-year period. With the rich biological and
behavior history of the rat model and the upcoming genomic tools and genetic
applications, a repository for the large number of current, and future, inbred,
transgenic, and congenic strains is a high priority.
4. Interaction with the NIH Mouse Mutagenesis and
Phenotyping Program ($1 million per mouse center)
Mutagenesis and phenotyping of rat models, in parallel
with NIH mouse mutagenesis and phenotyping program will allow significant
interaction and collaboration on developing new rodent models of human disease,
with comparable characterization and shared expertise.
IV. BENEFITS
The rat is a principal model organism to link function
to genes. The biological relevance and wealth of phenotypic data in the rat,
when combined with the current and proposed genomic resources, will provide the
opportunity to develop new diagnostic, prevention, and treatment approaches for
human health and disease.
REPORT OF THE NIH RAT MODEL
PRIORITY MEETING
I. BACKGROUND
The rat model provides important strengths for the
study of human health and disease. The large number of inbred rat models and
the vast amount of data (physiological, behavioral, biochemical, cellular,
pharmacological, toxicological, etc.) provide a superb platform on which to
build the genetic and genomic tools and resources to delineate the connections
between genes and biology. Importantly, in many instances, the rat is the most
appropriate experimental model of human disease.
The NIH, realizing the potential of rat models in
understanding basic biology and human health and disease, launched the Rat
Genome Program in 1995, followed by the Rat EST Program in 1997. These two
programs, funded by 13 Institutes and Centers at NIH, have produced a variety
of basic genomic resources. In addition, these programs have begun to provide a
powerful tool to link and capitalize upon the data and resources of other model
organisms and the human.
An era of extraordinary opportunities and
unprecedented scientific discovery now presents itself. New developments in
genomics, genetics, drug discovery, stem cell research, bioengineering, and
other fields are creating opportunities for revolutionary changes in the
practice of medicine. To facilitate the translation of these discoveries to
improved human health, the rat model, which remains a dominant biological
discovery tool, must be poised to take advantage of the opportunities
available, with suitable investments to capitalize on these opportunities.
As a consequence, the National Institutes of Health
(NIH) Director, Dr. Harold Varmus, asked the National Heart, Lung, and Blood
Institute (NHLBI) to convene a meeting to discuss the opportunities and
prioritize the needs to fully take advantage of rat models. The NHLBI, in
conjunction with 18 NIH Institutes and Centers, organized a broad-based meeting
of distinguished scientists to identify needs and opportunities, establish
priorities, and recommend costs.
II. STRENGTHS
The rat is a principal model organism to link function
to genes. The biological relevance and wealth of phenotypic data in the rat,
when combined with the current and proposed genomic resources, provide the
opportunity to accelerate the development of new diagnostic, prevention, and
treatment approaches for human health and disease.
A. BIOLOGY
The rat model has made enormous contributions to our
present understanding of biological function and behavior. The rat has been a
widely studied model system, as demonstrated by the number of publications in
the last three decades (nearly 500,000 PubMed publications). Large numbers of
rat disease models exist (more than 250 inbred, congenic, mutant, or transgenic
rat strains) to explore disease-related variables. Modeling of human diseases
can capitalize on the considerable strengths that these rat models offer to the
future of physiological and functional genomics, and in delineating genes of
complex diseases. Many of the rat models have already proven their utility for
addressing the human condition. Presently rats comprise 28% of laboratory
animals (AALAC) and provide important models for cardiovascular, pulmonary,
renal, endocrinology, reproduction, toxicology, parasitology, immunology,
development of dental plaque and gingivitis, polycystic kidney disease,
spongioform encephalopathy, alcoholism, nutrition, cancer, growth, diabetes,
autoimmune disease, arthritis, asthma, endocrinology, multiple sclerosis,
learning, memory, behavior, and neurological health and disease. In some cases,
specific aspects of human disease are recapitulated well only in the rat,
making these animals a unique resource for studying and identifying genetic
pathways relevant human disease. Many examples exist of biological relevance to
human health and disease, and several were presented at the meeting and/or
discussed on the Rat Community Forum (http://goliath.ifrc.mcw.edu/RCF/, this
Web page is no longer available) prior to the meeting. Some examples
include:
Cardiac Function and Hypertension: The
rat is a model of choice for many physiological studies related to cardiac and
vascular function, pulmonary circulation, energetics and metabolism,
microcirculation, neural control of cardiovascular, renal and pulmonary
function, age and gender related differences, studies of arterial pressure
regulation, hypertension, cell and system integrative function, and signal
transduction studies. Many inbred rat strains are currently available and well
characterized (there are 9 inbred strains for arterial pressure regulation and
hypertension alone). An example of the strength of combining physiology and
genetics was described. Results were shown in which 220 phenotypes were
determined in a large F2 cross between an inbred strain of Dahl S and Brown
Norway rats in which multiple cosegregation analyses were performed using 220
"informative markers" that distinguish these two strains. More than 30 regions
on 16 different chromosomes were significantly related to measured parameters
that are likely determinants of blood pressure. Combining the results from
several rat strain crosses and examining the overlapping QTL regions allowed
the prediction of the location of several human chromosomal regions that had
been previously identified in blood pressure linkage analysis studies in human
populations. These results now enable investigators to develop models that
share phenotypic similarities to the clinical picture as well as share
homologous genomic regions responsible for the similarities, thereby providing
unprecedented opportunities for generating new models directly relevant to
human disease.
Behavioral and Neuropharmacology of
Addiction: There is an appreciable depth of knowledge of rat
neuroanatomy and neurophysiology. Complex behavioral procedures involved with
drug self-administration and developmental studies related to substance abuse,
including the behavioral effects of maternal drug exposure, have been
extensively characterized in the rat model. Three levels of biological analysis
used in the neurological study of alcohol and drug addiction (Intra-Cellular B
signal transduction processes; Trans-Cellular B signal transmission processes;
Multi-Cellular B signal integration processes) all use the rat successfully to
model human biology. Rat studies using models of ethanol self-administration
are providing important insight as to how alcohol and aggression interact, with
data that appear more related to the human situation than the other models
systems. One of the most important attempts to understand the lasting effects
of drug actions in the brain has been to look for permanent changes in brain
function following chronic drug administration, such as the effects in neural
sensitivity and function that last after long periods of drug abstinence. These
changes are believed central to the issues of drug taking relapse. That drug
and alcohol exposure has the potential for epigenetic effects altering gene
expression is the basis for several hypotheses as to the mechanism for these
lasting effects. These cellular studies in the rat have become the cornerstone
for this approach where connections are being developed between cellular
function and behavioral phenotype in the rat related to relapse. Gene by
environment interactions is a critical area for study in the addiction field.
The environmental exposure (light, noise, proximity, etc.) can affect the
interpretation of the genetic contribution to several of alcohols
behavioral effects. For instance, environmental factors have major effects on
ethanol preference in both heterogeneous and selected rat strains. While this
is an understudied area in all model organisms, the studies in rats using
complex behavioral self-administration procedures have a high importance, given
the availability of the several selected alcohol-preferring rat lines that meet
the criteria as a model of human alcoholism.
Arthritis and Related Autoimmune
Disorders: Rat models of arthritis and related autoimmune diseases are
biologically relevant models to common human diseases such as rheumatoid
arthritis, insulin-dependent diabetes, multiple sclerosis, and autoimmune
uveitis. More than 200 inbred (e.g., LEW, DA, BB-DP, BB-DR, F344, BN, ACI),
congenic (e.g., MHC and other loci), mutant (e.g., athymic nude), or transgenic
(e.g., HLA-B27, TNF-alpha, HTLV-1 env-pX) rat strains exist in which to explore
disease-related variables. Several important models of adjuvant and bacterial
cell wall arthritis are only available in the rat, as rats are naturally more
susceptible to these disease models. In addition, disease penetrance in mice
(as noted in the necessity for repeated injection of potent "adjuvants" for
disease induction) is usually less than observed in rats, complicating genetic
analyses. Likewise, there are several unique infectious arthritis models
available in rat (e.g., Yersinia enterocolitica and
Chlamydia trachomatis arthritis). There are unique examples
of gene by environment interactions in the rat (induction of insulin-dependent
diabetes in BB-DR rats, and induction of arthritis with low potency,
non-immunogenic adjuvants in DA rats) as well as responses to therapeutic
agents in rat (rats are responsive to non-steroidal anti-inflammatory drugs,
whereas mice are resistant).
Gender-related disease susceptibility profiles in rat
are similar to those observed in humans. Female rats are more susceptible (as
are humans) to most of the arthritis models than are males. In contrast, male
mice are more susceptible than females.
Learning, Memory, and Behavior: The past
100 years of behavioral research using the rat has revealed the complexity of
learning and memory, as well as the multiplicity of brain systems that support
it. These studies show that a combination of thorough behavioral
characterization and neurobiological investigations can provide major insights
into the specific brain systems that mediate memory. Continuing efforts that
respect the psychobiological character of rats have recently allowed
investigations of even more complex cognitive and memory capacities. For
example, the rats superb learning abilities have been exploited using
odors as cues and foraging as a modality for behavioral expression. In this
format rats show exceedingly rapid learning of simple discrimination problems -
acquiring them typically in 1-2 trials and retaining the information for at
least several days. With this capacity in hand, rats have been trained using
the same methods on very complex problems such a Piagets transitive
inference task, a test solved by human children at about the age of 7. Rats
show robust transitivity and this capacity is fully dependent on the
hippocampus.
Endocrinology and Reproductive Biology:
There are various aspects of rat husbandry that provide attractive features for
reproductive physiological work; rat pregnancies are more size consistent
(compared to the mouse), rat cycling is relatively non-pheromonal (similar to
human), rats can be bred quickly after parturition, and rat brains show early
sexual dimorphism.
Respiratory and Pulmonary Biology: Many
models of lung disease use rat lungs and/or rat cells. There is a large body of
literature in the rat on the neurophysiologic structures, interventions and
cardiorespiratory monitoring that enable productive investigation in
understanding sleep and breathing. One significant advantage of the rat model
is the ability to perform lung function studies. In the rat, sleep, breathing,
and cardiac function measurements can be simultaneously recorded. The
availability of detailed neurofunctional information (in addition to a history
of behavior studies) provides an efficient transition from genes to complex
behaviors such as sleep. In addition, the rat model mimics many features of
human asthma and acute lung injury. Similar phenotypic measures can be
accomplished in the rat and human, and have not proven successful in other
model systems.
Toxicology and Pharmacology:
Pharmacogenetics is a major emerging research area. Not surprisingly the rat
remains a dominant model system for risk assessment of virtually all forms of
therapeutics and chemical substances. Insofar as current risk assessment
protocols require more than one species it is critical to continue to develop
the rat for risk assessment. For example, the acceptance of transgenic animals
for risk assessment linked to the increased availability of this technology in
rats provides for developing better models systems. Therefore, the combination
of classical risk assessment with genetic susceptibility to chemical agents
provides unparalleled opportunities for linking the vast databases on drug
responses to the genome, as well as increasing our understanding of gene-drug
interactions. Cancer: The rat models for breast cancer are good
representations of human breast cancer. They are hormonally responsive, can be
rapidly induced in virus free animals, and their histopathology and
premalignant stages of development resemble those of human breast cancer. The
great majority of cancer chemoprevention models in use today are rat based.
B. PHENOTYPING
One of the major strengths of the rat model is the in
depth characterization and well defined, relevant phenotypic measures. The size
of the rat allows many important measures to be quantified, without a delay
caused by needing to develop new technologies, including: invasive procedures
(intravenous cannulation for drug administration or blood collection, surgical
manipulations, nerve recordings, blood pressure monitoring, etc.), chronic
measurements (regional blood flows, cardiac output, etc.), collection of
tissues (synovium, lymph nodes, pituitaries, retina, heart, etc.), and serial
blood collections. In addition, arthritis of individual joints in rats can be
precisely described in terms of the day of onset, the pattern of onset, the
number and distribution of joints involved, the severity of swelling or
inflammation (in mice, since the joints are so small, descriptions are usually
limited to gross descriptions of swelling of whole digits or entire feet). The
general metabolic rate in mouse is approximately 3 times that of the rat and,
therefore, issues of the duration of drug action and dose are a problem when
studying addiction processes in the mouse. The absolute dimensions of the rat
has advantages for some studies, such as the use of multiple electrodes and
transducers, or the injection of neuroanatomical markers, where lack of
resolution through diffusion seen in a smaller brain is not a problem in the
rat. All of these size issues are exacerbated in developmental studies, both
prenatal and neonatal. As miniaturization of techniques, assays, and equipment
occur, careful attention needs to be paid to the increase in skill and
particularities needed, as otherwise reproducibility of data may suffer.
Technical challenges will need to be met in order to obtain conscious
measurements of cardiac output and regional blood flow, to carry out CNS
recordings and infusion, and to miniaturize all of the many biochemical assays.
However, many of these are not insurmountable and, by working together, the
rodent model users can find ways to accomplish these goals.
The field of cardiovascular physiology has seen a
major animal model change in the 1970s, when it became increasingly difficult
to use the mongrel dog. The transition from the dog to the rat as the
predominant research model in cardiovascular research occurred slowly over
about a 15 year period as retooling related to size and scaling factors
occurred and young investigators were trained in requisite new techniques.
Although there are relatively fewer investigators doing systems physiology now,
as compared to the 1970s and 1980s, with an appropriate investment of time and
resources, a solid understanding of mouse physiology and biochemistry could,
and should, be obtained. However, as stated previously, the rat model most
closely mimics the human condition for many health and diseases areas, because
of a different basic biology that cannot be recapitulated by ENU-mutagenesis in
the mouse. Therefore, investment in the genomic and genetic tools for rat will
be a sound and cost-effective approach for understanding pathobiology and
developing new diagnostic, prevention, and treatment approaches for human
health and disease.
III. ACCOMPLISHMENTS, NEEDS, AND
OPPORTUNITIES
The workshop participants identified two general area
of opportunities and needs. These two areas are biology/physiology and genomic
infrastructure. Biology/physiology needs and opportunities consisted of
germ-line modification, a National Rat Genetic Resource Center, a Rat Genome
Database, mutagenesis and phenotyping, and strengthening the rat model user
research community. Genomic infrastructure needs and opportunities consisted of
an expanded EST program, SNP development and mapping, enhanced BAC libraries,
and pilot DNA sequencing. Each opportunity and need is described below.
A. BIOLOGY/PHYSIOLOGY
1. Germ-line Modification
The need for access to the germ-line of the rat in
order to produce informative mutants is critical. The availability of gain of
function (over-expression; knock-in) and loss of function (knock-out) mutations
in the rat will be necessary for the functional characterization of genes.
Correct assignment of quantitative trait loci (QTL) to genes will require
verification from loss of function mutations or gene replacement studies.
Although the production of transgenic rats is now routine, the creation of loss
of function mutations by homologous recombination in embryonic stem (ES) cells
has not yet been possible in the rat. The production of ES like cells for the
rat has been accomplished in a number of labs using both standard methodology
or selective ablation of differentiated cells with Oct4 promoters in transgenic
blastocysts. Although these cells are morphologically like ES cells and carry a
variety of markers that indicate they have not committed to differentiation
pathways and hence are likely to be pluripotent, they have so far failed to
produce germ-line chimeras useful for generation of targeted mutations.
An alternative approach for routine production of loss
of function mutations in a variety of strains and transgenics is nuclear
transfer (NT), in which a nucleus from cultured cells with targeted mutations
is placed in the enucleated egg of the animal and then developed to term. The
nucleus so transferred carries the mutation desired and, for all practical
purposes, defines the strain of the offspring obtained. Animals produced by
nuclear transfer of embryonic or adult cells have been obtained from primates,
pigs, rabbits, mouse, cows, and sheep. Offspring produced from zygotic fusion
have been obtained in the rat. Animals produced by nuclear transfer with
genetically modified cells have been obtained for cows and sheep, suggesting
the general strengths of the overall strategy.
Nuclear transfer consists of several steps, including
egg obtainment, enucleation of the egg, fusion of the donor cell or insertion
of the donor nucleus, activation of the egg and transfer to a surrogate mother
for development to term. In the rat, many labs have achieved the efficient
collection of eggs by superovulation with hormones, laying the ground-work for
pursing studies in this area. Both fusion and injection of nuclei lead to
preimplantation development in the rat. Development of live offspring following
transfer of embryos to surrogate mothers is routine in the rat. The birth of an
offspring from this process would provide a heterozygous mutant animal if the
donor nucleus had come from a cell with a heterozygous targeted mutation.
The efficiency of embryo transfer with respect to
development of live births is greatly influenced by time spent in suboptimal
culture. Moreover, the assessment of developmental potential of the eggs
following NT is best done using in vitro culture allowing direct visualization
of the preimplantation stages of development. However, there are very few
systems that allow robust development from one cell fertile eggs to the
blastocyst stage of the rat embryo. One such system, R1ECM, works well for
Wistar outbred and Sprague Dawley outbred rats, but not for other, particularly
inbred, strains. Progress with NT would be accelerated if a culture system that
worked well with many inbred as well as outbred strains were available.
2. National Rat Genetic Resource Center
In August 1998, NIH convened a meeting entitled NIH
Rat Model Repository Workshop (http://www.nhlbi.nih.gov/nhlbi/sciinf/model/ratmodel.htm).
The recommendation of the workshop was, in order to meet the needs of the broad
rat research community and to provide the foundation for consistent and
well-characterized rat models for human disease, to establish a national,
central repository resourceCa National Rat Genetic Resource Center (NRGRC). The
main functions of the NRGRC would be as follows: (1) maintain and
characterize the most widely used rat strains; (2) preserve valuable
strains, including transgenic strains, through cryopreservation; (3)
distribute genetically and microbiologically high-quality animals;
(4) provide information, advice, and training in the use of genetically
defined rat strains; (5) contribute to the research and development
of technological advances in cryopreservation, embryo culture, and animal
maintenance; (6) serve as a platform for scientific discourse and
international cooperation among the community of scientists utilizing the rat
as a model system by sponsoring workshops and annual symposia.
The participants of this current meeting unanimously
agreed that the NRGRC is central to effective rat model research and endorsed
the recommendations of the previous report. With the rich biological and
behavior history of the rat model and the upcoming genomic tools and genetic
applications, a repository for the large number of current, and future, inbred,
transgenic, and congenic strains is a high priority.
3. Rat Genome Database
In response to the recommendations of the Rat Genome
Advisory Committee and the Report of the NIH Model Organism Database Workshop
(http://www.nhlbi.nih.gov/), the NIH
has issued a request for applications (RFA) for a Rat Genome Database (RGD).
The objective of this RFA is to establish a database that will collect,
consolidate, and integrate data generated from ongoing rat genetic, genomic,
and related research efforts, and to make these data widely available to the
scientific community. The applications were due on April 30, 1999 and an award
is expected September 30, 1999. The participants strongly endorsed this plan
for NIH to implement an RGD. Rat genomic, genetic, and phenotypic data needs to
be easily and readily accessible to all researchers. The participants also
requested that the Rat Community Forum web site continue, although support for
the web site was not discussed. Since the meeting participants considered the
implementation of the RGD to be underway, it was not considered in the final
priority list. However, the RGD was seen as absolutely essential.
4. Interaction with the NIH Mouse Mutagenesis and
Phenotyping Program
Mutagenesis and phenotyping of rat models, in parallel
with the NIH mouse mutagenesis and phenotyping program, will allow significant
interaction and collaboration on developing new rodent models of human disease,
with comparable characterization. Random mutagenesis can offer the rat model an
alternative to gene targeting strategies, as many of the induced mutations are
loss of function. In addition, if rat and mouse mutagenesis and phenotyping
programs are established together, each can benefit from the others experience.
There is significant intellectual capital in the rat physiology and behavior
communities. This knowledge-base should be involved in the translation of the
physiological and behavioral methods needed by the mouse phenotyping program
and to establish comparative studies of these procedures in the mouse and rat
models.
5. Strengthening the Rat Model User Research
Community
A major success of the meeting was catalyzing the
formation of an interactive research community of rat-users. A web site (Rat
Community Forum) was established in advance of the meeting to pose a series of
8 questions to this community at large. The community was identified via email
lists from several societies affiliated with FASEB. The member names were
screened against Medline and rat-users identified. The rat-users were then sent
a single message to visit a web site to comment on the 8 questions posed for
the meeting. In total more than 130 rat-users from a diverse range of fields
responded to the questions. These responses, in conjunction with the diversity
of research topics represented by the meeting participants, provided a unique
opportunity to exchange ideas about building unity, capitalizing on
opportunities, and exchanging ideas within this community. Dr. Varmus suggested
the community develop a rat contact group to interface with the NIH. Several
suggestions were made to further develop the community:
a. The Rat Community Forum (RCF) web site
(http://goliath.ifrc.mcw.edu/RCF/, this Web page is no longer available)
should remain open.
b. Participation in a series of rat genetic based
meetings scheduled.
1. Physiological Genomics in the Rat, Cold Spring
Harbor Laboratories. December 59, 1999.
2. Rat Genetics and Genomics Meeting, Goteborg,
Sweden. June 13-16, 2000.
3. Rat Genetic and Genomics Meeting, Milwaukee,
WI. Summer, 2001.
c. Satellite rat genetics meeting in conjunction
with The 13th International Mammalian (Mouse) Genetics Conference,
October 31November 3, 1999 in Philadelphia, PA., as well as future
International Mammalian Genome Society activities.
d. Satellite rat genetics and phenotyping meeting in
conjunction with the American Society of Human Genetics.
Many of these meetings and interactions are already
being planned. The participants were very enthusiastic about continued
interaction, with NIH providing support through staff involvement (identify
speakers and topics, develop agendas, etc.) and funds for conference grants and
meetings.
B. GENOMIC INFRASTRUCTURE
Recognizing the usefulness of the rat as a model
system, NIH funded the Rat Genome Project (RGP) and the Rat Expressed Sequence
Tag (REST) Project to develop important genomic tools and resources that will
further enhance the power of rat model systems (Table 1). These two
infrastructure projects, and the RFA for the Rat Genome Database, provide the
ability to link the rat physiology and functional biology with the genetic
tools of the mouse and the clinical features of the human through comparative
mapping to facilitate the translation of bench to bedside. However, much is
left to be done to fully and effectively capitalize on the opportunities the
rat provides.
Table 1: Existing Rat Genomic
Tools
Tool |
Site(s) Where Developed |
Genetic Markers and genetic maps |
Whitehead/MIT, MGH, MCW, University of Iowa,
NIH, Oxford and Otsuka Pharmaceutical Company |
YAC libraries |
Whitehead/MIT, German Rat Genome
Project |
PAC library |
Roswell Park Cancer Institute |
BAC library |
Roswell Park Cancer Institute |
Radiation Hybrid Panel |
Cambridge/Research Genetics |
Radiation Hybrid Map |
MCW, University of Iowa, Oxford and Otsuka
Pharmaceutical Company |
Normalized cDNA libraries |
University of Iowa |
I.M.A.G.E clones |
U of Iowa, Research Genetics |
Rat ESTs |
U of Iowa, TIGR |
Rat UniGene Clusters |
NCBI |
Dense Mapping Cross |
German Genome Project, MCW |
1. Rat EST (REST) Project
The REST project has generated more than 93,000 ESTs
derived from 12 different normalized cDNA libraries. These normalized libraries
combined with a serial subtraction strategy have provided an unprecedented
level of efficiency with respect to identifying unique rat genes, a set of
27,000 UniGene clusters. This is a marked improvement over the 15,000 mouse
UniGene clusters generated from more than 300,000 ESTs. The REST also has the
goal of mapping 8,000 ESTs on the radiation hybrid map. Additionally, 1,500
ESTs with sequence homology to human ESTs or genes have also been placed on the
RH map, thereby facilitating the development of more accurate comparative maps.
The NIH has approved a competitive renewal of the REST. If the applications are
approved, they would increase the number of UniGene clusters in the rat to
60,000, with 27,000 of these ESTs mapped onto the radiation hybrid (RH) map.
The participants were very enthusiastic about the progress of this project, and
recommended the goal be expanded to achieve 90% coverage of all rat genes. It
was recognized that this goal would require careful monitoring to insure that
novel genes were continuing to be discovered at reasonable cost.
2. SNP Discovery and Mapping
The rat genetic map and the rat radiation hybrid map
have more than 8,000 genetic markers (primarily microsatellites). These maps
enable investigators to identify chromosomal regions (QTLs) that contain genes
responsible for specific phenotypes. It is evident from both this meeting and
the literature that a large number of investigators are using rats to
successfully locate QTLs that are models for common human disease. However, the
density of markers currently available is nearly an order of magnitude short of
the number of markers that will be required to positionally clone genes from
the mapped positions. To overcome this limitation, the participants suggested
constructing a 3rd generation genetic map using single nucleotide polymorphisms
(SNPs) at a resolution of 1 SNP per 100 kb.
3. BAC Clone Resources
One of the major goals of the RGP is to build the
initial physical mapping tools in the form of a series of large insert
libraries for genomic DNA. The International Rat Genome effort has generated
two 10-fold rat YAC libraries, a >10-fold PAC library, and a >10 fold BAC
library. These physical mapping reagents are useful for initiating individual
positional cloning projects and are being used by many laboratories around the
world. However, with the change in sequencing capacity in both the public and
the private sector it is important to consider if these reagents position the
rat for the sequencing of its full genome, once the mouse genome is sequenced.
Several of the participants have evaluated the BAC resources and determined
that the size of the inserts combined with the use of a single restriction
enzyme limit the use of existing BAC resources for sequencing. Therefore, there
is a need to develop a BAC library with 15-fold coverage made from multiple
sources of DNA cut using a variety of restriction enzymes or random shear
techniques while still maintaining an average insert size greater than 150
kilobases.
4. Genomic Sequence
Within less than two years the sequencing capacity of
the publicly funded US and International Genomics community is likely to pass
that needed to sequence a mammalian genome to 10-fold redundancy in one year.
The rat should be positioned for genomic sequencing immediately after, or in
parallel, with the mouse. Support for near term finished sequencing of 5-10
megabases of the rat genome in at least 1 megabase parcels should be a goal.
Syntenic regions corresponding to mouse and human genomic intervals slated for
immediate sequencing should be prioritized. Detailed analysis of this sequence,
community response, as well as simulations based on this data set, will provide
an empiric basis for selecting an appropriate genomic sequencing strategy for
the rat.
II. RECOMMENDATIONS AND PRIORITIES
The needs and opportunities for the rat model
identified and described above were prioritized at the meeting by the
participants. Although the initial discussions grouped the recommendations into
two groups (biology/physiology and genomic infrastructure), the prioritizing
discussions pulled out the most critical initiatives based on need and
opportunity.
The final recommendations are listed in priority
order.
1. Germ-line Modifications ($8.5 million
over 5 years)
(See Accomplishments, Needs, and Opportunities;
Biology/Physiology; Germ-line Modifications: section III/A/1)
Germ-line modification in the rat is critically
important to assigning gene function to specific genes and to identifying gene
alterations responsible for specific phenotypes. The analysis of phenotypes
from gain of function and loss of function has been the most direct and useful
way to connect specific genes to phenotypes relevant to human disease. Although
the production of transgenic rats is routine, the accessibility to this
technology is limited and needs to be extended. Transportation and storage of
valuable transgenics is problematic. The development of ES like cells in the
rat is routine, but these fail to make germ-line chimeras and therefore cannot
be used to take advantage of homologous recombination for knock-outs and
knock-ins. The highest priority should be to overcome these limitations,
thorough the use of the following strategies: (a) The development of nuclear
transfer in the rat is an important priority that needs to be met ($5 million
over 5 years). (b) The development of dominant negative mutations by pronuclear
injection should be encouraged ($500,000 per year for 5 years). (c)
Cryopreservation of zygotes and sperm will help to alleviate the storage and
transportation problems ($750,000 over 5 years). (d) The generation of in vitro
fertilization (IVF) techniques in the rat, including intracytoplasmic sperm
injection (ICSI), will help the use of cryopreserved sperm by allowing
suboptimal sperm to lead to viable offspring ($250,000 over 2 years). ICSI has
been achieved in the rat and its use could be easily optimized and made
generally available. The developments from this research could be transferred
to the NRGRC for implementation in that resource and for distribution to the
research community.
2. Additional Genomic Resources ($30
million over 3 years)
(See Accomplishments, Needs, and Opportunities;
Genomic Infrastructure: section III/B/1,2, 3,4)
a. EST Project Enhancement ($2.5 million per year
for 3 years): The participants were very enthusiastic about the progress of
this project, and recommended that the goal be expanded to achieve 90% coverage
of all rat genes. Achieving this goal will offer significant advantages and
opportunities. It will identify rare transcripts in specific tissues of
interest, as well as in specific developmental stages. It will also allow the
development of a finer syntenic map with the mouse and human, which will
greatly improve efforts in identifying genes and elucidating their function.
b. SNP Discovery and Mapping ($6.5 million over 3
years): Developing a SNP map of the rat is a high priority, as it is required
to accelerate the identification of genes (through positional cloning)
responsible for complex, common diseases. There are multiple strategies for
developing SNPs. While no formal requirement was stated as to how SNPs should
be developed, the participants recommended that the existing rat EST project be
used to leverage NIH resources. As outlined previously, one of the goals of the
competitive renewal of REST (assuming funding) is to develop a rat UniGene set
consisting of 60,000 ESTs. Of these 60,000 ESTs, 27,000 will be mapped on the
RH map and therefore provide ideal starting points for the SNP map, as the
primers that have been developed for the mapping can be used for SNP selection.
The other 33,000 ESTs also provide a rich source of sequence data for
initiating the development of SNPs, assuming this project is renewed
(competitive) this year.
c. BAC Clone Resources ($1 million for 1 year):
Sequencing the rat genome should be placed at very high priority as the third
mammalian genome to be sequenced after man and mouse. However, the necessary
clone resources must be developed quickly. The capacity of the publicly funded
genome sequencing efforts will most likely exceed 20 raw megabases per year by
the spring of 2000. It therefore seems likely that aggressive sequencing of the
rat could commence before 2004, with a high quality draft being possible a year
thereafter. The key prerequisite for such an effort is a high quality well
characterized, and deeply redundant BAC library. This library should have large
inserts (average size greater than 150 kilobases), be redundant to a depth of
15-fold or greater, and ideally be produced by more than one restriction enzyme
or physical shearing. The currently available BAC library is unsatisfactory
with respect to insert size, and is the product of a single restriction
enzyme.
Additionally, at least a 10-fold subset of these BAC
clones should be subjected to end sequencing and restriction fingerprinting.
This is an essential resource to permit efficient genomic sequencing with a
minimum investment in mapping.
d. Pilot Sequencing ($15 million over 3
years): A complete sequence of the rat genome will provide the critical
substrate for understanding the molecular basis of biological function and
pathology. A coordinated and systematic approach will be most cost effective.
It is however not yet clear how to optimally sequence the rat given the
existence of the sequence for mouse and man, a rich array UniGene models from
all three species, and the need to address further genomes. It may be that some
form of draft sequence in the rat would strike the best
cost/benefit balance. To address this question, a small number of rat genome
sequencing pilot studies, each covering a few megabases, should be performed.
Regions homologous to ones already sequenced in both man and mouse should be
selected. The already available rat BAC library should be used for the source
of clones.
3. National Rat Genetic Resource Center
($35 million over 5 years)
(See Accomplishments, Needs, and Opportunities;
Biology/Physiology; National Rat Genetic Resource Center: section III/A/2)
The genetic integrity of, and access to, important rat
strains is maintained by an international effort that depends on the personal
good will of many individuals. This effort is inherently inefficient and
susceptible to the vicissitudes of funding and local interests. A much more
robust approach to the problem of genetic integrity, microbiological quality
and distribution of these valuable models would be the National Rat Genetic
Resource Center (NRGRC).
The objectives of the NRGRC are to serve as a
national, central resource that will select, maintain, distribute, and preserve
genetically defined rats; to coordinate the extramural NRGRC activities with
the intramural NIH Genetic Resource; to develop a cost-effective central
resource that will maintain the maximum number of strains without compromising
the quality of strains; to establish criteria of strain selection,
preservation, and distribution of genetically defined rats to the research and
supplier communities; to facilitate and implement the establishment of
standards for genetic, phenotypic, and microbiological monitoring; to
disseminate information concerning germ-line modification techniques to the
scientific community; to provide relevant information to the scientific
community via a Web page that interfaces with other rat databases; and to
sponsor meetings to discuss various uses of the rat in biomedical research and
the developments in rat genetics and genomics.
Establishment of the NRGRC will have a broad impact on
a wide range of research areas by providing an effective source of quality,
transportable animals and embryos that will meet the current needs and
anticipated increased demand due to the development of important genomic tools
and resources in the rat. Lack of accessibility to strains of known
microbiological and genetic quality is a major limitation to studies using
inbred rat models, as commercial suppliers carry a very small subset of inbred
rat strains and genetic purity data are not always available.
If necessary, a smaller pilot of the NRGRC could be
envisioned to establish utility. The participants noted that the cost of the
NRGRC could be reduced by adopting user fees and encouraged NIH to consider
creative solutions to the long-term support of the repository, including a
potential commercial supplier to fill this critical niche.
4. Interaction with the NIH Mouse Mutagenesis and
Phenotyping Program ($1 million per mouse center)
(See Accomplishments, Needs, and Opportunities;
Biology/Physiology; Interaction with the Mouse Mutagenesis and Phenotyping
Program: section III/A/4)
Mutagenesis and phenotyping of rat models should be
incorporated into the NIH Mouse Mutagenesis and Phenotyping RFA program. Direct
interactions between the rat physiologists and the mouse geneticists will
enhance the utility of discovering and characterizing new models for human
disease. Direct participation will also increase the likelihood of technology
and know-how transfer between the rat and mouse research communities. In this
way, the mouse research community can gain expertise and knowledge in
phenotypic methods, while the rat research community will gain expertise and
knowledge in genetic manipulation. The involvement of the rat physiologists and
behaviorists in translating established norms and protocols used in the rat to
be applied in the mouse will be of great benefit to both rat and mouse user
communities.
This direct collaboration and translation will allow
empirical data to be gathered using side by side comparisons of each model in
various physiological, biochemical, behavioral, and surgical measures and
procedures. In addition, genome-wide, random mutagenesis offers an alternative
approach to gene targeting strategies.
V. SUMMARY
Rat models offer exciting opportunities to understand
human health and disease. Their use to understand complex diseases and
biological phenomena requires the highest quality and most effective genomic
and genetic tools and resources and a functional infrastructure. Such
comprehensive resources, as delineated in this report, are needed for effective
identification of genes responsible for disease and health, defining gene
function, understanding how genes interact with the environment and with each
other, discovering and testing new drugs, and designing new prevention
strategies.
NIH Rat Model
Priority Meeting Lawton Chiles International Center
Bethesda, MD May 3, 1999 Agenda |
Plenary
Session |
7:15 a.m. |
Registration and Continental Breakfast
|
|
8:00 a.m. |
Introduction |
Dr. Varmus |
8:20 a.m. |
Overview |
Dr. Branscomb
Dr. Cowley |
8:30 a.m. |
Rat Genome Tools, Resources, and
Applications |
Dr. Jacob |
8:55 a.m. |
Cardiovascular Physiology |
Dr. Cowley |
9:20 a.m. |
Behavioral and Neuropharmacology of
Addiction |
Dr. Samson |
9:45 a.m. |
Break |
|
10:10 a.m. |
Arthritis and Related Autoimmune
Disorders |
Dr. Wilder |
10:35 a.m. |
Learning, Memory, and Behavior |
Dr. Eichenbaum |
11:00 a.m. |
Reproductive Physiology and
Endocrinology |
Dr. Conn |
11:25 a.m. |
Rat Models Using Transgenesis |
Dr. Mullins |
11:45 a.m. |
Embryonic Stem Cells and Nuclear Transfer
Technologies in the Rat |
Dr. Iannaccone |
12:05 p.m. |
Follow Up Discussions |
Dr. Branscomb Dr. Cowley |
12:30 p.m. |
Lunch |
|
1:30 p.m. |
Issue A: Comparative Value |
Dr. LaVail |
|
Question 2: What are the
strengths and weaknesses of rat models of health and disease, and how do they
compare to the strengths and weaknesses of mouse models (for example, what can
be done in the rat that cant be done in the mouse; what can be done in
the mouse that cant be done in the rat)?
Question 3: Are the rat and
mouse sufficiently close evolutionary and physiologically to each other that
continued investment should be restricted to one mammalian model system?
Question 5: Does the rat occupy
a unique and essential place in studying health and disease that cannot be
served by technological (e.g. miniaturization of assay systems) advances in the
mouse? |
2:00 p.m. |
Issue B: Functional Genomics |
Dr. Walker |
|
Question 1: What is the likely impact of these
genomic resources and reagents for the rat on defining gene function and
increasing our understanding of common diseases that afflict humans? |
2:30 p.m. |
Issue C: Infrastructure |
Dr. Blankenhorn |
|
Question 4: How would further
development of genetic and genomic infrastructure for the rat (e.g., ES cells,
nuclear transfer, Jackson Lab-like repository for the rat, synteny
map(human/mouse/rat), genomic sequence, database) enhance the utility of the
Human Genome Project and our ability to understand common human diseases? What
will be lost if the rat infrastructure does not keep pace with other model
organisms and the Human Genome Project?
Question 8: What are the
consequences of not developing the infrastructure of the rat any further?
|
3:00 p.m. |
Break |
|
3:30 p.m. |
Issue D: Physical Resources |
Dr. Duyk |
|
Question 6: Within less than two
years the sequencing capacity of the publicly funded US and International
Genomics Community is likely to pass that needed to sequence a mammal to 10X
redundancy in one year. The second major target for this capacity will
certainly be the mouse. What impact should these expectations have on how
genomic infrastructure should be developed for the rat. What is the optimal
targeting approach for sequencing in the rat given such capacities and the
existence of full genomic sequence from mouse and human? What resources should
be made available, and when to allow rat genomic sequencing to be most
effectively undertaken?
Question 7: Are there benefits
of having mouse, human, and rat genomic sequence available? |
4:00 p.m. |
Needs, Opportunities, and Future Directions
|
Dr. Branscomb
Dr. Cowley |
5:00 p.m. |
Adjourn |
|
ATTENDEE ROSTER
CO-CHAIRPERSONS
Elbert W. Branscomb, Ph.D. Director DOE
Joint Genome Institute Building 100 2800 Mitchell Drive Walnut
Creek, CA 94598 (925) 296-5608 (925) 296-5710 FAX branscomb1@llnl.gov
Allen W. Cowley, Ph.D. Professor and Chairman
Department of Physiology Medical College of Wisconsin 8701
Watertown Plank Road Milwaukee, WI 53226 (414) 456-8277 (414)
456-6546 FAX cowley@mcw.edu
MEMBERS
Kenneth M. Baldwin, Ph.D. Professor
Physiology and Biophysics NASA Life Sciences Physiology and
Biophysics U.C. Irvine Irvine, CA 92697 (949) 824-7192 (949)
824-8540 FAX kmbaldwi@uci.edu
J. Carl Barrett, Ph.D. Scientific Director
Division of Intramural Research National Institute of Environmental Health
Sciences National Institutes of Health P.O. Box 12233 111 T.W.
Alexander Drive Research Triangle Park, NC 27709 (919) 541-3205
(919) 541-7784 FAX barrett@niehs.nih.gov
Elizabeth P. Blankenhorn, Ph.D. Professor
Department of Microbiology and Immunology MCP Hahnemann University
2900 Queen Lane Philadelphia, PA 19085 (215) 991-8392 (215)
848-2271 FAX libby@mcphu.edu
Kristina Borror, Ph.D. Science Policy Analyst
Office of Science Policy National Institutes of Health Building
1, Room 218 9000 Rockville Pike Bethesda, MD 20892 (301)
496-6836 (301) 402-0280 FAX borrork@od.nih.gov
R. Nick Bryan, M.D., Ph.D. Director of Diagnostic
Radiology Clinical Center Building 10, Room 1C660 Bethesda, MD
20892 (301) 435-5741 (301) 496-9933 FAX
nbryan@mail.cc.nih.gov
William L. Castleman, D.V.M., Ph.D. Professor of
Pathology Department of Pathobiology University of Florida,
Gainesville P.O. Box 110880 2015 Southwest 16th Avenue
Gainesville, FL 32610 (352) 392-4700 EXT 3920 (352) 392-9704 FAX
mailto:cnbryan@mail.cc.nih.gov
P. Michael Conn, Ph.D. Oregon Health Sciences
University Oregon Regional Primate Research Center 505 Northwest
185th Avenue Beaverton, OR 97006 (503) 690-5297 (503) 690-5569
FAX connm@ohsu.edu
Christopher P. Cunningham, Ph.D. Professor and
Interim Chair Department of Behavioral Neuroscience Oregon Health
Sciences University 3181 Southwest Sam Jackson Park Road Portland, OR
97201-3098 (503) 494-2018 (503) 494-6877 FAX
cunningh@ohsu.edu
Geoffrey Duyk, M.D., Ph.D. Chief Scientific
Officer Exelixis Pharmaceuticals, Inc. 260 Littlefield Avenue
South San Francisco, CA 94080 (650) 825-2232 (650) 825-2205 FAX
duyk@exelixis.com
Howard Eichenbaum, Ph.D. Professor
Department of Psychology Boston University 64 Cummington Street
Boston, MA 02215 (617) 353-1426 (617) 353-1414 FAX
hbe@play.bu.edu
Gerald Fischbach, M.D. Director Office of
the Director National Institute of Neurological Disorders and Stroke
National Institutes of Health Building 31, Room 8A52 Bethesda,
MD 20892-9663 (301) 496-9746 (301) 496-0296 FAX gf33n@nih.gov
Janina R. Galler, M.D. Director Center for
Behavioral Development and Mental Retardation Boston University School of
Medicine M921B 85 East Newton Street Boston, MA 02118 (617)
638-4840 (617) 638-4843 FAX Jgaller@bu.edu
Michael Garrick, Ph.D. Department of Biochemistry
and Pediatrics SUNY at Buffalo 140 Farber Hall 3435 Main Street
Buffalo, NY 14214-3000 (716) 829-3926 (716) 829-2725 FAX
mgarrick@buffalo.edu
Michael Gould, Ph.D. Professor
Oncology/McArdle Labortory University of Wisconsin Room K4/332
600 Highland Avenue Madison, WI 53792 (608) 263-6615 (608)
263-9947 FAX gould@humonc.wisc.edu
Joan T. Harmon, Ph.D. Senior Advisor for Diabetes
Division of Diabetes, Endocrinology, and Metabolic Diseases National
Institute of Diabetes and Digestive and Kidney Diseases National
Institutes of Health 45 Center Drive Bethesda, MD 20892 (301)
594-8813 (301) 480-3503 FAX joan_harmon@nih.gov
Richard J. Hodes, M.D. Director National
Institute on Aging National Institutes of Health Room 5C35 31
Center Drive Bethesda, MD 20892 (301) 496-9265 (301) 496-2525
FAX hodesr@31.nia.nih.gov
Steven E. Hyman, M.D. Director National
Institute of Mental Health National Institutes of Health Room 8235
6001 Executive Boulevard Bethesda, MD 20892-9669 (301) 443-3673
(301) 443-2578 FAX shyman@nih.gov
Christine E. Kasper, Ph.D., R.N., F.A.A.N.,
F.A.C.S.M. M. Adelaide Nutting Chair School of Nursing Johns
Hopkins University Room 432 525 North Wolfe Street Baltimore, MD
21205 (410) 502-5402 (410) 955-4890 FAX
ckasper@son.jhmi.edu
Stephen I. Katz, M.D., Ph.D. Director
National Institute of Arthritis and Musculoskeletal and Skin Diseases
National Institutes of Health Building 31, Room 4C32 31 Center Drive
Bethesda, MD 20892-2350 (301) 496-4353 (301) 480-6069 FAX
katzs@niams.nih.gov
A.B. Kulkarni, Ph.D. Senior Investigator
National Institute of Dental and Craniofacial Research National Institutes
of Health 30 Convent Street Bethesda, MD 20892 (301) 435-2887
(301) 435-2888 FAX ak40m@nih.gov
Matthew LaVail, Ph.D. Professor Anatomy and
Ophthalmology University of California, San Francisco Room K120
10 Kirkham Street San Francisco, CA 94143-0730 (415) 476-4233
(415) 476-0709 FAX mmlv@itsa.ucsf.edu
Theresa Lee, Ph.D. Program Director
Molecular Biology Division of Basic Reseach National Institute on
Drug Abuse National Institutes of Health Room 4282, MSC 9555
6001 Executive Boulevard Bethesda, MD 20892-9555 (301) 443-6300
(301) 594-6043 FAX tl37h@nih.gov
Claude Lenfant, M.D. Director Office of the
Director National Heart, Lung, and Blood Institute National
Institutes of Health Building 31, Room 5A-52 31 Center Drive MSC 2486
Bethesda, MD 20892-2486 (301) 496-5166 (301) 402-0818 FAX
lenfantc@gwgate.nhlbi.nih.gov
Jon Levine, M.D., Ph.D. Professor Department
of Oral and Maxillofacial Surgery University of California, San Francisco
Campus Box 0440 521 Parnassus Avenue, Room C522 San Francisco,
CA 94143-0440 (415) 476-5108 (415) 476-6305 FAX
levine@itsa.ucsf.edu
Yvonne Maddox, Ph.D. Deputy Director Office
of the Director National Institute of Child Health and Human Development
National Institutes of Health Building 31, Room 2A03 Bethesda,
MD 20892 (301) 496-1848 (301) 402-1104 FAX
maddoxy@exchange.nih.gov
Robert J. Mason, M.D. Professor Department
of Medicine National Jewish Medical and Research Center 1400 Jackson
Street Denver, CO 80206-1997 (303) 398-1302 (303) 398-1806 FAX
masonb@njc.org
John P. Mordes, M.D. Professor of Medicine
Diabetes Division Department of Medicine University of Massachusetts
Medical School Biotech II, Suite 218 373 Plantation Street
Worcester, MA 01605 (508) 856-3800 (508) 856-4093 FAX
john.mordes@umassmed.edu
John Mullins, Ph.D. Professor Centre for
Genome Research University of Edinburgh Kings Building
West Mains Road United Kingdom +44-131-650-5864 +44-131-667-0164
FAX j.mullins@ed.ac.uk
Joe Nadeau, Ph.D. Professor Department of
Genetics Case Western Reserve University BRB 630 10900 Euclid
Avenue Cleveland, OH 44106-0581 (216) 368-0581 (216) 368-3432
FAX jhn4@po.cwru.edu
Allan I. Pack, M.D., Ph.D. Professor of Medicine
Center for Sleep Department of Medicine Hospital of the
University of Pennsylvania 991 Maloney Building 3600 Spruce Street
Philadelphia, PA 19104-4283 (215) 662-3302 (215) 662-7749 FAX
pack@mail.med.upenn.edu
Daniel Rotrosen, M.D. Director Division of
Allergy, Immunology, and Transplantation National Institute of Allergy and
Infectious Diseases National Institutes of Health Solar Building,
Room 4A24 Bethesda, MD 20892 (301) 496-1886 (301) 402-2571 FAX
drotrosen@niaid.nih.gov
Norka Ruiz Bravo, Ph.D. Acting Director
Division of Cancer, Biology National Cancer Institute National
Institutes of Health MSC 7380 6130 Executive Boulevard
Rockville, MD 20892-7380 (301) 435-5225 (301) 496-8656 FAX
nb96@nih.gov
Herman H. Samson, Ph.D. Professor Department
of Physiology and Pharmacology School of Medicine Wake Forest
University Medical Center Boulevard Winston Salem, NC 27157-1083
(919) 716-8590 (919) 716-8501 FAX hsamson@wfubmc.edu
Martin Sarter, Ph.D. Professor Department of
Psychology Ohio State University Townshend Hall, Room 27 1885
Neil Avenue Columbus, OH 43210 (614) 292-1751 (614) 688-4733 FAX
sarter.2@osu.edu
David W. Self, Ph.D. Assistant Professor
Department of Psychiatry Yale University 34 Park Street New
Haven, CT 06508 (203) 974-7727 (203) 974-7897 FAX
david.self@yale.edu
Judy A. Small, Ph.D. Chief Division of
Extramural Research CAIB National Institute of Dental and
Craniofacial Research National Institutes of Health Building 45, Room
4AN-24J Bethesda, MD 20892 (301) 594-2425 (301) 480-8318 FAX
judy.small@nih.gov
Judith L. Vaitukaitis, M.D. Director
National Center for Research Resources National Institutes of Health
Building 31, Room 3B11 31 Center Drive Bethesda, MD 20892-2128
(301) 496-4793 (301) 402-0006 FAX
vaitukaitis@nih.gov
Harold E. Varmus, M.D. Director National
Institutes of Health Building One, Room 126 One Center Drive MSC 0148
Bethesda, MD 20892-0148 (301) 496-2433 (301) 402-2700 FAX
Cheryl L. Walker, Ph.D. Professor Science
Park-Research Division Department of Carcinogenesis M.D. Anderson
Cancer Center University of Texas P.O. Box 389 Park Road 1C
Smithville, TX 78957 (512) 237-9550 (512) 237-2475 FAX
cwalker@odin.mdacc.tmc.edu
Thomas J. Wronski, Ph.D. Professor
Department of Physiological Sciences College of Veterinary Medicine
University of Florida, Gainesville P.O. Box 100144, JHMHC 1600
Southwest Archer Road Micanopy, FL 32667 (352) 392-4700 EXT 3844
(352) 392-5145 FAX wronskit@mail.vetmed.ufl.edu
Annette Wysocki Scientific Director National
Institute of Nursing Research National Institutes of Health MSC 0967
Building 9, Room 1W125 Bethesda, MD 20892-3583 (301) 402-3583
(301) 435-3435 FAX awysocki@box-a.nih.gov
NIH PLANNING
COMMITTEE
Peter A. Dudley, Ph.D. Program Director
Division of Extramural Research (DER) National Eye Institute National
Institutes of Health Suite 350 6120 Executive Boulevard
Bethesda, MD 20892 (301) 496-0484 (301) 402-0528 FAX
pad@nei.nih.gov
Stephen L. Foote, Ph.D. Acting Division Director
Division of Neuroscience and Behavioral Science National
Institute of Mental Health National Institutes of Health Room 11-103
6001 Executive Boulevard, MSC 8030 Bethesda, MD 20892 (301)
443-1576 (301) 443-4822 FAX sfoote@helix.nih.gov
Robert W. Karp, Ph.D. Program Director
Department of Genetics National Institute on Alcohol Abuse and Alcoholism
National Institutes of Health Suite 402 6000 Executive
Boulevard, MSC 7003 Bethesda, MD 20892-7003 (301) 443-2239 (301)
594-0673 FAX rkarp@willco.niaaa.nih.gov
Steven Klein, Ph.D. Health Scientist
Administrator Development Biology, Genetics and Teratology National
Institute of Child Health and Human Development National Institutes of
Health 6100 Executive Boulevard Bethesda, MD 20892 (301)
496-5541 (301) 402-0303 FAX sk56d@nih.gov
Gabrielle LeBlanc, Ph.D. Health Scientist
Administrator Fundamental Neuroscience and Developmental Disorders
National Institute of Neurological Disorders and Stroke National
Institutes of Health Federal Building 916 7550 Wisconsin Avenue
Bethesda, MD 20892 (301) 496-5745 (301) 402-1501 FAX
leblancg@nswide.ninds.nih.gov
Donna R. Maglott, Ph.D. National Center for
Biotechnology Information National Library of Medicine National
Institutes of Health Building 38A, Room B2N14 Bethesda, MD 20892
(301) 435-5895 (301) 480-9241 FAX
maglott@ncbi.nlm.nih.gov
Cheryl L. Marks, Ph.D. Program Director
Cancer Genetics Branch, DCB National Cancer Institute National
Institutes of Health Room 5010 Executive Plaza North 6130
Executive Boulevard Bethesda, MD 20892-7381 (301) 435-5226 (301)
496-8656 FAX cm74v@nih.gov
Catherine McKeon, Ph.D. Senior Advisor for
Genetic Research National Institute of Diabetes and Digestive and
Kidney Diseases National Institutes of Health Building 45, Room
5AN-18B Bethesda, MD 20892-6600 (301) 594-8810 (301) 480-3503
FAX mckeonc@ep.niddk.nih.gov
Stephen C. Mockrin, Ph.D. Acting Director
Division of Heart and Vascular Diseases National Heart, Lung, and Blood
Institute National Institutes of Health Rockledge II, Room 9170
6701 Rockledge Drive, MSC 7940 Bethesda, MD 20892 (301) 435-0477
(301) 480-1336 FAX mockrins@gwgate.nhlbi.nih.gov
Nancy Nadon, Ph.D. Head Office of Biological
Resources National Institute on Aging National Institutes of Health
GW 2C231 7201 Wisconsin Avenue Bethesda, MD 20892 (301)
496-6402 (301) 402-0010 FAX nadonn@exmur.nia.nih.gov
Susan E. Old, Ph.D. Health Scientist
Administrator Division of Heart and Vascular Diseases National Heart,
Lung, and Blood Institute National Institutes of Health Rockledge II,
Suite 9150 6701 Rockledge Drive, MSC 7940 Bethesda, MD 20892-7940
(301) 435-0477 (301) 480-1336 FAX
olds@gwgate.nhlbi.nih.gov
Nancy J. Pearson, Ph.D. Chief Genetic
Sciences Initial Review Group Center for Scientific Review National
Institutes of Health Room 6178 6701 Rockledge Drive, MSC 7890
Bethesda, MD 20892-7890 (301) 435-1047 (301) 480-2067 FAX
pearsonn@csr.nih.gov
Jane L. Peterson, Ph.D. Program Director
National Human Genome Research Institute National Institutes of Health
Building 38A, Room 614 38 Library Drive Bethesda, MD 20892
(301) 946-2338 (301) 480-2770 FAX
jane_peterson@nih.gov
Gregory Schuler, Ph.D. Staff Scientist
National Center for Biotechnology Information National Library of Medicine
National Institutes of Health 8600 Rockville Pike Bethesda, MD
20894 (301) 435-7226 (301) 435-7794 FAX
schuler@ncbi.hlm.nih.gov
Hilary Sigmon, Ph.D., R.N. Physiologist, Nurse
Scientist Administrator National Institute of Nursing Research
National Institutes of Health Natcher Building, Room 5B10 31 Centers
Drive, MSC 2178 Bethesda, MD 20892-2178 (301) 594-5970 (301)
480-8260 FAX hsigmon@ep.ninr.nih.gov
Rochelle K. Small, Ph.D. Health Science
Administrator Division of Human Communication National Institute on
Deafness and Other Communication Disorders National Institutes of Health
Building EPS, Room 400 6120 Executive Boulevard, MSC 7180
Bethesda, MD 20892-7180 (301) 402-3464 (301) 402-6251 FAX
rochelle_small@nih.gov
John D. Strandberg, DVM., Ph.D. Director
Comparative Medicine National Center for Research Resources National
Institutes of Health Room 5146 One Rockledge Centre Bethesda, MD
20892-7965 (301) 435-0884 (301) 480-3558 FAX
johns@nerr.nih.gov
William A. Suk, Ph.D. Chief Division of
Extramural Research and Training Chemical Exposures and Molecular Biology
Branch National Institute of Environmental Health Sciences National
Institutes of Health P.O. Box 12233 Research Triangle Park, NC 27709
(919) 541-0797 (919) 541-2843 FAX
suk@niehs.nih.gov
Neal West, Ph.D. Health Science Administrator
Comparative Medicine National Center for Research Resources
National Institutes of Health Room 6166 6705 Rockledge Drive
Bethesda, MD 20892-7965 (301) 435-0744 (301) 480-3819 FAX
nealw@ep.ncrr.nih.gov
Ronald L. Wilder, M.D., Ph.D. Chief
Inflammatory Joint Diseases Section, ARB National Institute of Arthritis
and Musculoskeletal and Skin Diseases National Institutes of Health
Building 10, Room 9N240 9000 Rockville Pike Bethesda, MD 20892
(301) 496-6499 (301) 402-0012 FAX
wilderr@arb.niams.nih.gov
Paul B. Wolfe, Ph.D. Program Director
Division of Genetics and Developmental Biology National Institute of
General Medical Sciences National Institutes of Health Building 45
Bethesda, MD 20892 (301) 594-0943 (301) 480-2228 FAX
wolfep@nigms.nih.gov
EXECUTIVE COMMITTEE
Elbert W. Branscomb, Ph.D. Director DOE
Joint Genome Institute Building 100 2800 Mitchell Drive Walnut
Creek, CA 94598 (925) 296-5608 (925) 296-5710 FAX
branscomb1@llnl.gov
Allen W. Cowley, Ph.D. Professor and Chairman
Department of Physiology Medical College of Wisconsin 8701
Watertown Plank Road Milwaukee, WI 53226 (414) 456-8277 (414)
456-6546 FAX cowley@mcw.edu
Phillip M. Iannaccone, M.D., Ph.D. George M.
Eisenberg Professor Department of Pediatrics Northwestern University
Medical School and Childrens Memorial
Institute for Educational Research Mailstop 204 2300 Childrens
Plaza Chicago, IL 60614 (773) 880-8236 (773) 880-8266 FAX
pmi@nwu.edu
Howard J. Jacob, Ph.D. Associate Professor
Department of Physiology Medical College of Wisconsin 8701 Watertown
Plank Road Milwaukee, WI 53226 (414) 456-4887 (414) 456-6516 FAX
jacob@mcw.edu
Stephen C. Mockrin, Ph.D. Acting Director
Division of Heart and Vascular Diseases National Heart, Lung, and Blood
Institute National Institutes of Health Rockledge II, Room 9170
6701 Rockledge Drive, MSC 7940 Bethesda, MD 20892-7940 (301) 435-0477
(301) 480-1336 FAX mockrins@gwgate.nhlbi.nih.gov
Susan E. Old, Ph.D. Health Scientist
Administrator Division of Heart and Vascular Diseases National Heart,
Lung, and Blood Institute National Institutes of Health Rockledge II,
Suite 9150 6701 Rockledge Drive, MSC 7940 Bethesda, MD 20892-7940
(301) 435-0477 (301) 480-1336 FAX
olds@gwgate.nhlbi.nih.gov
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