From: kristoff@NET.BIO.NET (Dave Kristofferson)
Newsgroups: bionet.sci-resources
Subject: NIH Guide, vol. 21, no. 36, pt. 3, 9 October 1992
Date: 8 Oct 92 23:57:23 GMT
Distribution: bionet


$$XID RFA HL9303B HL-93-03-B P1O1 **************************************

CELL AND MOLECULAR BIOLOGY OF MEGAKARYOCYTOPOIESIS

NIH GUIDE, Volume 21, Number 36, October 9, 1992

RFA:  HL-93-03-B

P.T. 34; K.W. 0780015, 0780020, 0760003, 0760020, 1002008, 1002019

National Heart, Lung, and Blood Institute

Letter of Intent Receipt Date:  January 15, 1993
Application Receipt Date:  March 15, 1993

PURPOSE

The Division of Blood Diseases and Resources (DBDR), of the National
Heart, Lung, and Blood Institute (NHLBI) invites applications for
studies that may contribute to better understanding of regulation of
the proliferation and maturation of megakaryocytes and the control of
platelet production.  The knowledge gained may allow manipulation of
megakaryocytopoiesis and platelet level in patient populations.

HEALTHY PEOPLE 2000

The Public Health Service (PHS) is committed to achieving the health
promotion and disease prevention objectives of "Healthy People 2000,"
a PHS-led national activity for setting priority areas.  This RFA,
Cell and Molecular Biology of Megakaryocytopoiesis, is related to the
priority area of platelet disorders and bone marrow transplantation.
Potential applicants may obtain a copy of "Healthy People 2000" (Full
Report:  Stock No. 017-001-00474-0 or Summary Report:  Stock No.
017-001-00473-1) through the Superintendent of Documents, Government
Printing Office, Washington, DC 20402-9325 (telephone 202-782-3238).

ELIGIBILITY REQUIREMENTS

Applications may be submitted by domestic and foreign, for-profit and
non-profit organizations, public and private, such as universities,
colleges, hospitals, laboratories, units of State and local
governments, and eligible agencies of the Federal government.  Awards
in response to this RFA will be made to foreign institutions only for
research of very unusual merit, need, and promise, and in accordance
with PHS policy governing such awards.  Foreign institutions are not
eligible for First Independent Research Support and Transition
(FIRST) Awards (R29).  Applications from minority individuals and
women are encouraged.

MECHANISM OF SUPPORT

This RFA solicits applications for the National Institutes of Health
(NIH) individual research grant (R01) and FIRST Award (R29) and is a
one-time solicitation.  Applicants, who will plan and execute their
own research programs, are requested to furnish their own estimates
of the time required to achieve the objectives of the proposed
research project.  Up to five years of support may be requested.  At
the end of the official award period, renewal applications may be
submitted for peer review and competition for support through the
regular grant program of the NHLBI.  It is anticipated that support
for the present program will begin in September 1993.  Administrative
adjustments in project period and/or amount of support may be
required at the time of the award.  All current policies and
requirements that govern the research grant programs of the NIH will
apply to grants awarded in connection with this RFA.

FUNDS AVAILABLE

Although the financial plans for fiscal year 1994 include $1.5
million for this program, award of grants pursuant to this RFA is
contingent upon receipt of funds for this purpose.  It is anticipated
that about six grants will be awarded under this program.  The
specific amount to be funded will, however, depend on the merit and
scope of the applications received and on the availability of funds.
Since a variety of approaches would represent valid responses to this
announcement, it is anticipated that there will be a range of costs
among individual grants awarded.  If collaborative arrangements
involve sub-contracts with other institutions, the NHLBI Grants
Operations Branch (telephone 301-496-7257) may be consulted regarding
procedures to be followed.

RESEARCH OBJECTIVES

The bone marrow megakaryocytes are the cells that produce the blood
platelets.  The number of platelets and their biochemical integrity
are important for normal hemostasis, and both of these parameters are
dependent upon the normal development of the parent cells, the
megakaryocytes.  The megakaryocyte is the least understood
hematopoietic cell of the bone marrow.  It possesses features unlike
those of the other hematopoietic cells:  the megakaryocyte nuclei are
highly polyploid, and several thousand platelets are formed from a
remarkable process of synthesis and organization of the cytoplasmic
compartment of each megakaryocyte.  The factors which regulate the
process of endomitosis, the extent of polyploidy, and the
ultrastructural changes which occur in the nucleus during maturation
are not understood.  Likewise, the factors which regulate cytoplasmic
development are not known.  Specific growth and maturation factors
which trigger endomitosis and cytoplasmic maturation of
megakaryocytes are not yet defined, in contrast to the considerable
knowledge that has been obtained concerning factors specific for red
and white cells.  The in vivo location of platelet production needs
to be defined; although it is generally thought that the
megakaryocytes release their platelets in the marrow, it has also
been suggested that the megakaryocytes travel to the lung and release
platelets in the lung.  Optimal methods for assessing the rate of
production of platelets have not been established.  The feedback
mechanisms by which thrombocytopenia signals an increase in
production of platelets have not been characterized.  Thus
considerable work remains to be done in understanding the
developmental regulation of megakaryocytes and the factors which lead
to platelet release.

Disorders of megakaryocytopoiesis can lead to serious clinical
problems.  The loss of normal regulation of the production of
platelets and abnormal platelet function are seen in
myeloproliferative disorders and leukemias.  In myeloproliferative
disorders, this can result paradoxically in hemorrhage in the
presence of thrombocytosis.  It is reasonable to suspect that the
biochemical development of the megakaryocyte is abnormal in these
circumstances.  However, no consistent abnormalities have been
defined in studies of proteins of platelets obtained from patients
with these disorders.  An understanding of the biochemical
abnormalities which cause the thrombopathies would be very important,
and would depend on definition of abnormalities in platelets and on
an improved understanding of factors that influence megakaryocyte
biochemistry.

A rapidly growing clinical area in which the understanding of
megakaryocytopoiesis is critical is bone marrow transplantation,
since patients undergoing transplantation suffer from a prolonged
loss of platelet production.  Approximately 15 percent of bone marrow
transplant recipients die due to primary or secondary hemorrhagic
complications, mainly due to thrombocytopenia.  On the average, each
patient requires about 100 to 150 units of platelet transfusion until
recovery of marrow function occurs, usually after 40 to 50 days.
Some patients remain thrombocytopenic for a prolonged period and may
become refractory to platelet transfusion therapy.  An understanding
of megakaryocytopoiesis and platelet production is likely to be
beneficial for the support of bone marrow recipients.  If appropriate
growth factors could be isolated and cloned, they will find ready
application not only in these patients but also in other cases where
thrombocytopenia is a concern, e.g., patients with AIDS or cancer.

Similarly, although probably by a different mechanism, sepsis in
neonates is associated with prolonged thrombocytopenia.  Another
clinically relevant area which is poorly understood is the frequent
occurrence of megakaryoblastic leukemias in children with Down's
syndrome; this phenomenon is found as a transient leukemia in
neonates and often develops into a full-blown leukemia later in
childhood.  The characterization of the genetics and biochemistry of
this disorder may be very useful in elucidating the process of
megakaryocytopoiesis.  It is important to develop an understanding of
growth factors and other factors influencing hematopoiesis in order
to maintain normal blood cell production and thus be able to improve
treatment for the bleeding problems.

A major area of interest has been the interaction of proliferation
and growth factors with megakaryoblasts and megakaryocytes, both at
the progenitor level and at the level of cytoplasmic maturation.
Thus far, this work has largely depended on a mixed population of
progenitor cells for the starting assay, and a major obstacle to
understanding the effects of growth factors has been the inability to
isolate a pure normal megakaryoblastic cell line.  Studies at the
single cell level or with a pure cell population with newer
techniques (e.g., cell sorter or polymerase chain reaction) could be
productive.  Several cell lines derived from megakaryocytic leukemias
have been described recently, some of which contain a fraction of
cells capable of achieving a ploidy of 8N.  These cells may be useful
in elucidating some of the early stages of polyploidy.  However, they
all appear to have multiple trisomies and chromosomal rearrangements
and deletions, and the contribution of these factors to the lack of
normal development is not understood.  The chromosomal abnormalities
may restrict the usefulness of these cell lines in understanding the
normal dynamics of both development of polyploidy and cytoplasmic
maturation.  The availability of megakaryocytic cell lines which
behave in a normal manner will significantly help to answer some of
the outstanding questions in this area.  These questions include
identification of lineage-specific factors involved in stem cell
commitment to the megakaryocytic lineage, nuclear transcription
factors which may be unique to megakaryocytes, factors which
stimulate DNA and protein synthesis in megakaryocytes, and factors
which lead to cytoplasmic maturation and finally to platelet
production from the mature megakaryocyte cytoplasm.

New technology, such as the use of transgenic animals, provides the
potential for manipulating megakaryocytopoiesis in vivo.  For
example, a portion of the regulatory region of the gene for a
platelet-specific protein, platelet factor 4 (PF4) was used to insert
foreign genes into the germ cells of mice and to obtain
megakaryocyte-specific expression of the foreign proteins.  The
transgene expression occurred earliest in the 2N megakaryocytes, the
construct was expressed in a developmentally accurate fashion, and
there were no major positional effects between two cell lines with
regard to transgene expression.  The selective expression of the
transgene in bone marrow progenitors and peripheral blood cells can
be employed to devise simple bone marrow assays to monitor the
transition and commitment of hemopoietic stell cells to megakaryocyte
precursors.  This may allow the identification, isolation and cloning
of the elusive megakaryocyte maturation and growth factors.  The
transgenic mouse can be utilized to selectively target gene products
to megakaryocytes and platelets and then study their effects in vivo.

Examples of Areas of Interest

The following are only examples and prospective applicants are urged
to use their own ideas as to the area of research.

Development of Cell Lines and Culture Systems

The critical molecular events which lead to commitment of progenitor
cells to the megakaryocytic lineage and to complete maturation of
megakaryocytes to platelet production have not been identified.
These studies require the development of cell lines which can enter
and complete the normal maturation cycle.  Immortalized cell lines
could be generated which can be triggered to undergo differentiation
under restricted conditions.  Another approach would be to develop
bone marrow culture systems which allow the transition of stem cells
to megakaryocytes to be specifically monitored.  This approach could
employ precursor cells isolated from transgenic mice that possess
megakaryocyte-specific promotors coupled to reporter molecules in
conjunction with purified hematopoietic growth factors.  The role of
known oncogenes, growth factor receptors, and other signaling
proteins could be investigated in the above systems with dominant
negative suppressor techniques, antisense approaches, or injection of
specific neutralizing antibodies.  The function of previously
unidentified components might be sought by somatic cell genetic
techniques coupled to expression cloning methods.

Nuclear and Cytoplasmic Development

The biochemical steps which are responsible for the development and
extent of polyploidy and for maturation of the cytoplasm and the
formation of granule and membrane systems are not defined.  The
molecules that are specific to megakaryocytes and to different stages
of development must be defined; new markers other than those
currently used in studies of platelets should be sought.  The cis
acting regions and transacting factors of genes specifically
expressed in megakaryocytes should be determined, and the effects of
hematopoietic growth factors and cytokines on these events should be
assessed.

Specific areas of study which might elucidate nuclear development
include the role of cyclins, kinases, phosphatases, and contractile
proteins in endomitosis.  Specific areas of study regarding
cytoplasmic development include stage-specific events in granule
formation and synthesis of membrane and cytoskeletal components,
including targeting of proteins to specific structures.  The
mechanism by which the cytoplasm becomes organized into the mature
platelets is unknown, and studies which could address this question
would be useful.  It would also be important to identify the specific
growth factors which control nuclear and cytoplasmic development and
platelet formation.

Sensors and Signals

Investigations are needed to clarify the sensors and molecular
signals which modulate the plasma levels of the megakaryocyte
specific growth factors and thereby regulate megakaryocytopoiesis.

Platelet Production

Newly available techniques might be used to clarify questions related
to platelet production.  For example, megakaryocytes that have been
genetically labeled with short-lived reporter molecules might serve
as in vivo markers for quantitating platelet production.  The site of
megakaryocyte fragmentation might be determined by injecting the
labeled cells and measuring the formation of the circulating
platelets.

Genetic Defects in Megakaryocytopoiesis

Chromosomal abnormalities have been associated with megakaryocytic
leukemias, notably in Down syndrome and in adult leukemias in
otherwise genetically normal individuals.  Other congenital syndromes
which include defects in megakaryocytopoiesis have been identified,
such as TAR syndrome and Epstein's syndrome.  In addition, a number
of animal models of defective megakaryocytopoiesis have been
described.  Characterization of the genetic abnormalities in these
various conditions could provide useful information on the factors
regulating megakaryocytopoiesis.

Disciplines and Expertise

Among the disciplines and expertise that may be appropriate for this
program are hematology, biochemistry, cell biology, medicine, and
molecular biology.

Exclusions

Epidemiological studies and large-scale clinical trials are
specifically excluded from this RFA.

SPECIAL REQUIREMENTS

Upon initiation of the program, the NHLBI will sponsor annual
meetings to encourage and exchange of information among investigators
who participate in this program.  In the preparation of the budget
for the grant application, applicants must request additional travel
funds for one meeting each year to be held in Bethesda, MD.
Applicants must also include a statement in the applications
indicating their willingness to participate in such meetings.

STUDY POPULATIONS

SPECIAL INSTRUCTIONS TO APPLICANTS REGARDING IMPLEMENTATION OF NIH
POLICIES CONCERNING INCLUSION OF WOMEN AND MINORITIES IN CLINICAL
RESEARCH STUDY POPULATIONS

NIH policy is that applicants for NIH clinical research grants and
cooperative agreements will be required to include minorities and
women in study populations so that research findings can be of
benefit to all persons at risk of the disease, disorder or condition
under study; special emphasis should be placed on the need for
inclusion of minorities and women in studies of diseases, disorders
and conditions which disproportionately affect them.  This policy is
intended to apply to males and females of all ages.  If women or
minorities are excluded or inadequately represented in clinical
research, particularly in proposed population-based studies, a clear
compelling rationale should be provided.

The composition of the proposed study population must be described in
terms of gender and racial/ethnic group.  In addition, gender and
racial/ethnic issues should be addressed in developing a research
design and sample size appropriate for the scientific objectives of
the study. This information should be included in the form PHS 398 in
Sections 1-4 of the Research Plan AND summarized in Section 5, Human
Subjects.  Applicants are urged to assess carefully the feasibility
of including the broadest possible representation of minority groups.
However, the NIH recognizes that it may not be feasible or
appropriate in all research projects to include representation of the
full array of United States racial/ethnic minority populations (i.e.,
Native Americans including American Indians or Alaskan Natives),
Asian/Pacific Islanders, Blacks, Hispanics).  The rationale for
studies on single minority population groups should be provided.

For the purpose of this policy, clinical research includes human
biomedical and behavioral studies of etiology, epidemiology, (and
preventive strategies), diagnosis, or treatment of diseases,
disorders or conditions, including but not limited to clinical
trials.

The usual NIH policies concerning research on human subjects also
apply.  Basic research or clinical studies in which human tissues
cannot be identified or linked to individuals are excluded.  However,
every effort should be made to include human tissues from women and
racial/ethnic minorities when it is important to apply the results of
the study broadly, and this should be addressed by applicants.

For foreign awards, the policy on inclusion of women applies fully;
since the definition of minority differs in other countries, the
applicant must discuss the relevance of research involving foreign
population groups to the United States' populations, including
minorities.

If the required information is not contained within the application,
the application will be returned.

Peer reviewers will address specifically whether the research plan in
the application conforms to these policies.  If the representation of
women or minorities in a study design is inadequate to answer the
scientific question(s) addressed AND the justification for the
selected study population is inadequate, it will be considered a
scientific weakness or deficiency in the study design and will be
reflected in assigning the priority score to the application.

All applications for clinical research submitted to NIH are required
to address these policies.  NIH funding components will not award
grants or cooperative agreements that do not comply with these
policies.

LETTER OF INTENT

The NHLBI requests that prospective applicants submit, by January 15,
1993, a letter of intent that includes a brief synopsis of the
proposed research and identification of any other participating
institutions.   Such letters are requested only for the purpose of
providing an indication of the number and scope of applications to be
received; therefore their receipt is usually not acknowledged.  A
letter of intent is not binding, and it will not enter into the
review of any application subsequently submitted, nor is it a
necessary requirement for publication.  This letter of intent is to
be sent to:

Dr. Charles L. Turbyfill
Division of Extramural Affairs
National Heart, Lung, and Blood Institute
Westwood Building, Room 553
Bethesda, MD  20892
Telephone:  (301) 496-7351
FAX:  (301) 496-7033

APPLICATION PROCEDURES

Applications are to be submitted on the research grant application
form PHS 398 (rev. 9/91).  This form is available in most applicant
institution's offices of sponsored research or business offices; and
from the Office of Grants Inquiries, Division of Research Grants,
National Institutes of Health, 5333 Westbard Avenue, Room 449,
Bethesda, MD 20892, telephone 301/496-7441.  Use the conventional
format for research-project grant applications and ensure that the
points identified in the Section on Review Considerations are
fulfilled.  FIRST Award applications must include at least three
sealed letters of reference attached to the face page of the original
application.  FIRST Award applications submitted without the required
number of reference letters will be considered incomplete and will be
returned without review.

To identify the application as a response to this RFA, check "YES" on
Item 2a of page 1 of the application and enter the title and number
of the RFA, Cell and Molecular Biology of Megakaryocytopoiesis, HL-
93-03-B.

The RFA label available in application form PHS 398 must be affixed
to the bottom of the face page of the original copy of the
application.  Failure to use this label could result in delayed
processing of your application.

Send or deliver the completed application and three signed, exact
photocopies of it to:

Division of Research Grants
National Institutes of Health
Westwood Building, Room 240
Bethesda, MD  20892**

At the same time, send two additional copies of the completed PHS 398
application to Dr. Charles Turbyfill at the address listed under
LETTER OF INTENT.  It is important to send these two copies at the
same time as the original and three copies are sent to the Division
of Research Grants, otherwise the NHLBI cannot guarantee that the
application will be reviewed in competition for this RFA.

Applications must be received by March 15, 1993.  An application
received after this date will be considered ineligible.

REVIEW CONSIDERATIONS

Upon receipt, applications will be reviewed for completeness by the
DRG and for responsiveness by the NHLBI.  Incomplete applications
will be returned to the applicant without further consideration.  If
the application is not responsive to the RFA, NHLBI staff will
contact the applicant to determine whether to return the application
to the applicant or submit it for review in competition with
unsolicited applications at the next review cycle.

Applications may be triaged by an NHLBI peer review group on the
basis of relative competitiveness.  The NHLBI will withdraw from
further competition those applications judged to be non-competitive
for award and notify the applicant Principal Investigator and
institutional official.  Those applications judged to be competitive
will undergo further scientific merit review.  Those applications
that are complete and responsive will be evaluated in accordance with
the criteria stated below for scientific/technical merit by an
appropriate peer review group convened by the NHLBI.  The second
level of review will be provided by the National Advisory NHLBI
Council/Board.

Review Criteria

The factors to be considered in the evaluation of scientific merit of
each application will be similar to those used in the review of
research grant applications, including the novelty, originality, and
feasibility of the approach; the training, experience and research
competence of the investigator(s); the adequacy of the experimental
design; the suitability of the facilities; and the appropriateness of
the requested budget to the work proposed.

The second level review will be by the National Heart, Lung, and
Blood Advisory Council.

AWARD CRITERIA

The anticipated date of award is September 1993.

Funding decisions will be made on the basis of scientific and
technical merit as determined by peer review, program needs and
balance, and the availability of funds.

Awards in response to this RFA will be made to foreign institutions
only for research of very unusual merit, need, and promise, and in
accordance with PHS policy governing such awards.

INQUIRIES

Inquiries regarding this request for applications to:

Dr. Pankaj Ganguly
Chief, Thrombosis and Hemostasis Branch
Division of Blood Diseases and Resources
National Heart, Lung, and Blood Institute
Federal Building, Room 5C14
Bethesda, MD  20892
Telephone:  (301) 402-2237
FAX:  (301) 402-1622

Fiscal and administrative matters to:

Ms. Jane R. Davis
Chief, Blood Division Grants Management Section
Division of Extramural Affairs
National Heart, Lung, and Blood Institute
Westwood Building, Room 4A15
Bethesda, MD  20892
Telephone:  (301) 496-7257
FAX:  (301) 402-1200

AUTHORITY AND REGULATIONS

The programs of the Division of Blood Diseases and Resources, NHLBI,
are described in the Catalog of Federal Domestic Assistance No.
93.839.  Awards will be made under the authority of the Public Health
Service Act, Section 301 (42 USC 241) and administered under PHS
grants policies and Federal Regulations 42 CFR Part 52 and 45 CFR
Part 74.  This program is not subject to the intergovernmental review
requirements of Executive Order 12372 or to Health Systems Agency
review.


$$XID RFA AI9208 AI-92-08 P1O1 *****************************************

PAPILLOMAVIRUS IN VITRO CELL CULTURE SYSTEMS

NIH GUIDE, Volume 21, Number 36, October 9, 1992

RFA:  AI-92-08

P.T. 34; K.W. 1002045, 0780015, 0740020

National Institute of Allergy and Infectious Diseases

Letter of Intent Receipt Date:  November 6, 1992
Application Receipt Date:  December 10, 1992

PURPOSE

The Antiviral Research Branch of the Division of Microbiology and
Infectious Diseases (DMID), National Institute of Allergy and
Infectious Diseases (NIAID), invites Cooperative Agreement
applications from organizational entities willing to participate,
with the assistance of the NIAID, in furthering innovative in vitro
approaches to the study of papillomavirus infections and their
therapeutic control.  The goals of this solicitation are to stimulate
the use of in vitro papillomavirus replication systems for research
on (1) the events of papillomavirus replication and pathogenesis and
(2) the antiviral potential of experimental therapeutic agents.

HEALTHY PEOPLE 2000

The Public Health Service (PHS) is committed to achieving the health
promotion and disease prevention objectives of "Healthy People 2000,"
a PHS-led national activity for setting priority areas.  This Request
for Applications (RFA), Papillomavirus In Vitro Cell Culture Systems,
is related to the priority area of sexually transmitted diseases.
Potential applicants may obtain a copy of "Healthy People 2000" (Full
Report:  Stock No. 017-001-00474-0) or 'Healthy People 2000" (Summary
Report:  Stock No. 017-001-00473-1) through the Superintendent of
Documents, Government Printing Office, Washington, DC 20402-9325
(telephone 202-783-3238).

ELIGIBILITY REQUIREMENTS

Applications may be submitted by domestic and foreign for-profit or
nonprofit organizations, public and private, such as universities,
medical colleges, hospitals, laboratories, units of State and local
governments, and eligible agencies of the Federal government having
adequate resources or access to resources to support papillomavirus
in vitro cell culture research.  Applications from minority
individuals and women are encouraged.

MECHANISM OF SUPPORT

Support for this program will be through the cooperative agreement
(U01) funding instrument.  The U01 is an assistance mechanism in
which NIAID programmatic involvement with the recipient during the
performance of the planned activity is anticipated.  The nature of
NIAID program involvement is described under SPECIAL REQUIREMENTS.
The awardee will be responsible for the planning, direction, and
execution of the proposed project and interrelated activities.
Except as otherwise stated in this RFA, the award will be
administered under PHS grants policy as stated in the Public Health
Service Grants Policy Statement, DHHS Publication No. (OASH)
90-50,000, revised October 1, 1991.  The total project period for
applications submitted in response to the present RFA may not exceed
four years.

This RFA is a one-time solicitation.  NIAID has no plans to reissue
this announcement at any future date.  Future competing continuation
applications will compete under the R01 mechanism with all
investigator-initiated applications and will be reviewed by an
appropriate DRG study section according to the customary peer review
procedures for unsolicited applications.

FUNDS AVAILABLE

The NIAID anticipates making at least one award as a result of this
request. However, the number of awards to be made is dependent upon
receipt of a sufficient number of applications of high scientific
merit and upon availability of funds.  Although this program is
provided for in the financial plans of NIAID, awards pursuant to this
RFA are contingent upon the availability of funds for this purpose.
If appropriate, collaboration with other investigators or
institutions is permissible.  It is expected that the total direct
costs for the first year for successful applications will be around
$175,000.  However, individual awards may be higher or lower.

In general, awards will be made for a project period of up to four
years.  The earliest possible award date is July 1, 1993.  Specific
selections for awards will be made by a NIAID/DMID senior staff
committee.  Criteria will be scientific excellence, expertise,
program balance, and cost.

RESEARCH OBJECTIVES

Background

Papillomavirus infections are the most rapidly spreading viral
sexually transmitted disease today.  It is estimated that as many as
24 million people in the United States are infected with HPV.  This
infection not only has a serious detrimental impact on the infected
individual's quality of life, but several types of the etiologic
agent are almost certainly a cofactor in the development of cervical
carcinoma which kills 7000 American women annually.  In
immunocompromised patients, warts may grow exuberantly and cause
serious morbidity.  In addition, recurrent respiratory papillomatosis
(RRP), a fortunately rare disease believed to be acquired during
passage through the birth canal of an infected mother, can be
life-threatening.

None of the currently available therapies, including interferon,
5-fluorouracil, cryotherapy, laser surgery and podophyllotoxin are
adequate.  Therefore, the NIAID made two awards in 1988 to
investigators to utilize rabbit, nude mouse, and primate models of
papillomavirus infections for the evaluation of experimental
therapies.  As a result of these studies, several compounds have been
identified which appear to warrant clinical evaluation.  Phase I
studies of one of these, ribavirin, as a therapy for RRP have
recently begun. Further preclinical study of the others, PMEG and
HPMPC, members of the phosphonate family of nucleotides, are targeted
for clinical studies in the near future.

Although preliminary evaluations of experimental agents are performed
in vitro prior to in vivo evaluation for all other DMID-supported
animal models, until recently there has not been a suitable in vitro
system for evaluation of anti- HPV agents.  In the past few years,
several research groups have developed in vitro systems in which many
of the events of papillomavirus replication have been reported to
occur.  The availability of these systems provides an unparalleled
opportunity to investigate the mechanisms of papillomavirus
replication and pathogenesis at a molecular level.  This basic
research may also result in the identification of viral genes which
are promising targets for therapeutic intervention.  In addition,
these systems should provide a means to evaluate the potential
anti-papillomavirus activity of experimental agents and provide a
rational basis for selecting agents whose activity warrants further
study in the animal models.

RESEARCH OBJECTIVES

The research to be supported by this RFA will be focussed on the use
of in vitro culture systems for studies of the multiple events of
virus replication including viral DNA replication and preferably
expression of late genes.  The culture systems will also be used to
study the effect of experimental therapeutic agents on these events.
Research applications should propose an in vitro system of studying
multiple replication events of papillomaviruses such as early gene
expression, viral DNA replication and preferably also the expression
of late genes.  Applicants should also provide a description of
research plans for study of compounds with therapeutic potential.

SPECIAL REQUIREMENTS

Terms and Conditions of Award

The Cooperative Agreement mechanism involves a partnership between
the recipient of the award and the NIAID.  The role of the NIAID is
to provide technical assistance and guidance to the Principal
Investigator.  The following terms and conditions pertaining to the
scope and nature of the interaction will be incorporated in the
Notice of Award.  These agreements will be in addition to the
customary programmatic and financial negotiations which occur in the
administration of grants.  The Terms of Award described in this
section are in addition to, and not in lieu of, otherwise applicable
OMB administrative guidelines; DHHS Grant Administration Regulations
at 45 CFR 74; other DHHS, PHS Grant Administration Policy Statement
and other NIAID administrative terms of award.

Decisions regarding a change of scope of the originally funded
application must be reviewed and approved by the NIAID Scientific
Coordinator in accord with the PHS Grants Policy Statement.  Such
changes may occur due to emerging research findings of other
investigators.  In the event of a dispute, the decision of a three
member arbitration team, described under Arbitration Procedures, will
be binding.

Although the NIAID Scientific Coordinator may participate in
discussions on research planning and data analysis, the recipient(s)
of these award(s) will be responsible entirely for the design,
conduct, and analysis of their research findings.  The NIAID
Scientific Coordinator will not direct, nor be liable for, the
awardee's research activities.  The awardee will be responsible for
publishing and disseminating research results.

Responsibilities of the Awardee

Under Cooperative Agreement assistance mechanisms, the NIH identifies
general or specific program areas for support, and the performers
define and implement the specific aims, objectives and approaches for
their awarded project activities. The Principal Investigator defines
the details for the project within the guidelines of the RFA, retains
primary responsibility for the performance of the scientific
activity, and agrees to accept close assistance of NIAID staff in the
aspects of scientific and technical management of the project as
specified in the Terms and Conditions of Award.

The data obtained will be the property of the awardee who will retain
custody of and primary rights to the data.

The awardee will be responsible for:

a.  Project Implementation and Management

The Principal Investigator is to define and implement the specific
aims, objectives, and approaches for in vitro research on
papillomaviruses.  Specifically, the awardee will be responsible for:

Defining research objectives and approaches in accord with his/her
own interests and perceptions of novel and feasible approaches to
conducting in vitro research on papillomavirus replication and the
effects of experimental antiviral agents on the events of
replication;

Designing, planning, and conducting the necessary research protocols
stipulated in the application;

Ensuring that the results obtained are analyzed, interpreted, and
published in a timely manner;

Modifying research protocols as appropriate; and Providing
information to the NIAID Scientific Coordinator concerning progress.

b.  Budget

The Principal Investigator should adequately justify the requested
budget for each twelve month segment.  Budget planning must include
such line items as personnel, major equipment, travel, etc.  Trips to
the NIAID must be projected once a year for the Principal
Investigator and possibly one or two other key personnel, if
appropriate.  Meetings will be for the purpose of maintaining project
cohesiveness, program and product standards, providing technical
assistance and other expertise.  The NIAID Scientific Coordinator
will attend each meeting.  Additional communications will be made by
telephone or telephone conference call.

c.  Reporting Requirements

An annual progress report is required.  In addition, the awardee will
keep the Scientific Coordinator apprised of experimental results on a
more frequent basis when the research involves the evaluation of an
experimental compound's anti-papillomavirus activity.

d.  Publications

The Government, via the Scientific Coordinator, will have access to
data generated under this cooperative agreement and may periodically
review the data and progress reports.  Information obtained from the
data may be used by the Scientific Coordinator for the preparation of
internal reports on the awardee(s)' activities.  However, the awardee
will retain rights to the data and timely publication is encouraged.
Publication or oral presentation of work done under this agreement is
the responsibility of the Principal Investigator and will require
appropriate acknowledgement of NIAID support.

Responsibilities of the NIAID

Assistance via U01s differs from the traditional research grant.  In
addition to the normal programmatic and administrative stewardship
responsibilities, the awarding component anticipates substantial
programmatic involvement during performance of the project.  Under
the U01 mechanism, NIAID staff will facilitate and participate in,
but not direct, the research in order to ensure that research
priorities for papillomaviruses are addressed.  A member of the NIAID
staff will serve as Scientific Coordinator and will participate as a
member of the research team.  The Scientific Coordinator will be the
Chief, Antiviral Research Branch, DMID.

The Scientific Coordinator will interact closely with the Principal
Investigator to assist with and facilitate the overall research
planning and data analysis.  During performance of the project the
NIAID Scientific Coordinator may provide assistance by participating
in the design of specific research group activities; advising in the
selection of sources or resources, replacement of staff; coordinating
or participating in collection and/or analysis of data; and, advising
in management and technical performance.

Involvement of the Scientific Coordinator comprises:

a.  Coordination between NIAID and Awardee.

The establishment of channels of collaboration and communication is
expected. Shortly after any award is made, key project personnel and
NIAID staff will meet to discuss provision of NIAID technical
assistance in accordance with the RFA guidelines.

b.  Collaborative Participation:

Providing assistance in avoiding unwarranted duplication of effort;

Retaining the option to suggest possible changes of direction in the
research endeavors based on the acquisition, and provision to the
awardee, of new information;

The NIAID Scientific Coordinator may suggest studies within the scope
of the award's objectives and research activities; may present to the
investigators experimental findings from published sources or from
contract projects in support of these suggestions; and may
participate in the design of experiments, and

Providing assistance, when appropriate, in designing protocols and in
analysis and interpretation of research data.

c.  Provision of Important Information and Collaborations:

Principal Investigators may not have immediate access to needed
information. Therefore, NIAID will facilitate meeting these needs and
ensure that the awardee(s) become active participants in the
Antiviral Research Branch Preclinical Research Program.  This Program
consists of:  (a) twelve U01 awards for research on potential
molecular targets for novel antivirals and strategies for the design
of new agents; (b) two N01 awards for the application of standard,
well-characterized assay methodologies for the evaluation of the
anti- herpesvirus and anti-respiratory virus potential of large
numbers of compounds; (c) seven N01 awards for animal models of human
viral infections (including papillomaviruses) that are used for
evaluation of experimental therapies and research on pathogenesis;
and (d) five U01 awards for studies of viral replication and
pathogenesis that are targeted towards the eventual development of
improved diagnostic methodologies.  The interaction of the awardee(s)
with other investigators in this program, as well as the consequent
early receipt of data from those laboratories pursuing related
research goals, will facilitate and expedite research progress.

d.  Provision of Needed Resources

The NIAID may provide assistance by suggesting potential sources of
agents for antiviral evaluation and/or facilitating the obtaining of
these agents by the Principal Investigator.  The specific agents to
be tested will be decided upon by the Principal Investigator with
assistance from the Scientific Coordinator. The application should
include an estimate of how many compounds with therapeutic potential
will be studied annually.

e.  Monitoring

There shall be periodic on-site or reverse-site monitoring of the
status of the Principal Investigator's research.  Such visits may
include discussions regarding scientific technique and methodology,
review of data, review of data analysis, suggestions to improve the
quality of the research, staff recruitment and training, and overall
evaluation efforts.  In addition, the NIAID Scientific Coordinator,
as a part of the annual review process, will review the status of the
research, using information obtained from site visits and the annual
progress reports to justify continued funding.

f.  Annual Meeting

The NIAID Scientific Coordinator will organize an annual symposium in
Bethesda, Maryland at which the Principal Investigator(s) will
discuss their progress.  This symposium will include other NIAID
Antiviral Research Branch-supported investigators engaged in
preclinical antiviral research.  This symposium will facilitate
evaluation of the feasibility of the attempted antiviral approaches,
and will promote productive interactions among Principal
Investigators.  Applicants should include funds for travel to this
meeting for the Principal Investigator, and other key personnel if
warranted, in their proposed budget.

g.  Ongoing Communication

In addition to the annual symposium, the NIAID Scientific Coordinator
will interact informally with each principal investigator through
additional meetings, averaging once a year, at the Principal
Investigator's institution or at professional scientific meetings as
well as by teleconferences, regular telephone calls, and written
communication.

Arbitration Procedures

The Terms and Conditions of Award, listed in this RFA, require that
the NIAID Scientific Coordinator participate in the decision-making
process concerning the continuation of an award based on successful
performance of the Awardee during the planning, developmental, and
implementation period of the U01.  Disagreements (e.g., programmatic,
technical, and evaluation) arising pursuant to these approvals will
be arbitrated by an Arbitration Panel composed of one award recipient
designee, one NIAID designee, and a third designee, with expertise in
the relevant area, chosen by the other two.  These special
arbitration procedures in no way affect the Awardee's right to appeal
an adverse action in accordance with PHS regulations at 42 CFR, Part
50, Subpart D and DHHS regulations at 45 CFR, Part 16.

LETTER OF INTENT

Prospective applicants are asked to submit, by November 6, 1992, a
letter of intent that includes a descriptive title of the proposed
research, and the names and affiliation(s) of proposed key
investigators.  The NIAID requests a letter of intent in order to
provide an indication of the number and scope of applications to be
received.  The letter of intent does not commit the sender to submit
an application, nor is it a requirement for submission of an
application.  The letter of intent will not enter into the review of
any application subsequently submitted.  It allows NIAID staff to
estimate the potential review workload and to avoid conflict of
interest in the review.

The letter of intent is to be sent to:

Dr. Olivia Preble
Chief, Microbiology and Immunology Review Section
Scientific Review Branch
National Institute of Allergy and Infectious Diseases
Solar Building, Room 4C20
6003 Executive Boulevard
Rockville, MD  20892

APPLICATION PROCEDURES

Applications are to be submitted on form PHS 398 (rev. 09/91).
Application kits are available at most institutional business
offices, and may be obtained from: Office of Grants Inquiries,
Division of Research Grants, National Institutes of Health, 5333
Westbard Avenue, Westwood Building, Room 449 Bethesda, MD 20892,
telephone (301) 496-7441.

The format and detail applicable to regular research grant
applications must be followed.

For purposes of identification and processing, mark "yes" in item 2a
on the face page of the application.  Also in item 2, under "number"
list the RFA number, which is RFA AI-92-08, and under title type in
"PAPILLOMAVIRUS IN VITRO CELL CULTURE SYSTEMS".  In item 2b, "type of
grant program", type in "COOPERATIVE AGREEMENT".  In addition, the
RFA label available in the application form PHS 398 must be affixed
to the bottom of the face page and placed on top of the entire
package.  Failure to use this label could result in delayed
processing of the application, such that it may not reach the review
committee in time for review.

The research proposed should describe plans to accommodate the RFA
research program requirements and NIAID staff involvement.

The original and three copies are to be mailed or delivered to:

Division of Research Grants
National Institutes of Health
Westwood Building, Room 240
Bethesda, MD  20892**

Two copies are to be mailed or delivered to:

Dr. Olivia Preble
Chief, Microbiology and Immunology Review Section
Scientific Review Branch
National Institute of Allergy and Infectious Diseases
Solar Building, Room 4C20
6003 Executive Boulevard
Rockville, MD  20892

Applicants using express mail or a courier service should change the
zip code to 20852 for the Solar Building.

Applications must be received by both DRG and Dr. Olivia Preble by
December 10, 1992.  Applications received after December 10, 1992
will be returned without review or will be sent to a DRG study
section as an unsolicited R01 for that study section's next review
cycle.  If the application submitted in response to this RFA is
substantially similar to a grant application already submitted to the
NIH for review, but has not yet been reviewed, the applicant will be
asked to withdraw either the pending application or the new one.
Simultaneous submission of identical applications will not be
allowed, nor will essentially identical applications be reviewed by
different review committees.  Therefore, an application cannot be
submitted in response to this RFA that is essentially identical to
one that has already been reviewed.  This does not preclude the
submission of substantial revisions of applications already reviewed,
but such applications must include an introduction addressing the
previous critique.

REVIEW CONSIDERATIONS

Applications will be received by the NIH Division of Research Grants
(DRG) and will usually be assigned to NIAID.  Should an application
contain research plans that significantly overlap with programs of
another institute, PHS Referral Guidelines will prevail in the
assignment of that application.

Review Procedures

Applications will be reviewed by DRG for completeness and by NIAID
staff to determine administrative and programmatic responsiveness to
this RFA.  Any application that is incomplete or does not meet the
minimum requirements of this RFA will be returned to the applicant
without scientific peer review.

Applications considered complete and responsive to the RFA may be
subjected to a triage review by an NIAID peer review group to
determine their scientific merit relative to the other applications
in response to the RFA.  The NIAID will withdraw from competition
those applications judged by the triage peer review group to be
noncompetitive for award and will notify the applicant Principal
Investigator and the institutional official.

Those applications judged to be competitive for award will be
reviewed by the standard NIH dual peer review system.  The initial,
or first level,  review for scientific and technical merit will be
conducted by the Scientific Review Branch of the Division of
Extramural Activities, NIAID during February/March, 1993.  The second
level of review will be provided by the National Advisory Allergy and
Infectious Diseases Council in June, 1993.

REVIEW CRITERIA

o  Factors to be considered in scientific evaluation of the
competitive applications include:

o  Originality and scientific merit of research approach, design, and
methodology as well as the potential scientific, technical, or
medical significance of the proposed research.  There will be special
interest in the advantages and limitations of proposed in vitro
culture systems.

o  Research experience and competence of the Principal Investigator
and staff to conduct the proposed studies.  The experience of the
Investigators with in vitro culture systems for papillomaviruses and
expertise in papillomavirus molecular biology are important.

o  Adequacy of time/effort which the Principal Investigator and staff
would devote to the proposed studies.

o  Adequacy of facilities and resources.

o  Reasonableness of proposed costs.

AWARD CRITERIA

While scientific merit is a prime consideration for selecting
applications for award, programmatic relevance, reasonableness of
proposed budget, and availability of funds also will be considered.

INQUIRIES

Direct inquiries relevant to the programmatic aspects of this RFA to:

Dr. Catherine Laughlin
Chief, Antiviral Research Branch
Division of Microbiology and Infectious Diseases
National Institute of Allergy and Infectious Diseases
Solar Building, Room 3A-22
6003 Executive Boulevard
Rockville, MD  20852
Telephone:  (301) 496-8285
FAX:  (301) 402-1456

Direct inquiries regarding review procedures and requirements to:

Dr. Olivia Preble
Chief, Microbiology and Immunology Review Section
Scientific Review Branch
National Institute of Allergy and Infectious Diseases
Solar Building, Room 4C-20
6003 Executive Boulevard
Rockville, MD  20852
Telephone:  (301) 496-8208
FAX:  (301) 402-2638

Direct inquiries regarding fiscal matters to:

Mr. Todd Ball
Grants Management Branch
National Institute of Allergy and Infectious Diseases
Solar Building, Room 4B-22
6003 Executive Boulevard
Rockville, MD  20852
Telephone:  (301) 496-7075
FAX:  (301) 480-3780

Timetable

Letter of Intent Receipt Date:     November 6, 1992
Application Receipt Date:          December 10, 1992
Review by Initial Review Group:    February/March 1993
Review by NIAID Advisory Council:  June 1993
Earliest Award Date:               July 1, 1993

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic
Assistance No. 93.856, Microbiology and Infectious Diseases Research.
Awards are made under authorization of the Public Health Service Act,
Title IV, Part A (Public Law 78-410; as amended.  Awards will be
administered under PHS grants policies and Federal Regulations 42 CFR
Part 52 and 45 CFR Part 74.  This program is not subject to the
intergovernmental review requirements of Executive Order 12372 or
Health Systems Agency review.