DEVELOPMENTAL PROCESSES IN DIFFERENTIAL EXPRESSION OF GLOBIN GENES

Release Date:  April 30, 1998

PA NUMBER:  PAS-98-060

P.T.

National Institute of Diabetes and Digestive and Kidney Diseases
National Heart, Lung, and Blood Institute

PURPOSE

The National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and
the National Heart, Lung and Blood Institute (NHLBI) invite investigators to
submit research grant applications to pursue basic investigations into the
developmental processes involved in the differential expression of globin genes. 
Applications from new investigators and investigators in training are
particularly encouraged.

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 PA, Developmental Processes In
Differential Expression Of Globin Genes, is related to the priority area of
chronic disabling diseases.  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 202-512-1800).

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.  Racial/ethnic minority individuals, women, and persons with
disabilities are encouraged to apply as principal investigators.

MECHANISM OF SUPPORT

Support of this program will be through the National Institutes of Health (NIH)
research project grant (R01) and exploratory/developmental grant (R21) support
mechanisms.

The direct costs per year for each R21 application must not exceed $75,000.  The
total project period for an R21 application funded in response to this PA may not
exceed two years and is not renewable.  The Exploratory/Developmental (R21)
research grant program provides limited funds for short-term research projects. 
These R21 grants provide an opportunity for initiating studies that may be
preliminary in nature.  Research investigators in relevant fields are invited to
apply for R21 grants in order to develop preliminary data that could form the
basis of future research project grant (R01) applications.

Applications from institutions that have a General Clinical Research Center
(GCRC) funded by the NIH National Center for Research Resources may wish to
identify the GCRC as a resource for conducting the proposed research.  If so, a
letter of agreement from either the GCRC program director or principal
investigator should be included with the application.

FUNDS AVAILABLE

A total of $2.0 million in total cost will be committed by each of the
participating Institutes to fund applications of high scientific merit submitted
in response to this Program Announcement.  Because the nature and scope of the
research proposed in response to this PA may vary, it is anticipated that the
size of an award will vary also; however, the support of requests exceeding the
NIH average size of $160,000 direct cost for R01 grants would be unusual and
would require ample justification.  Although this program is provided for in the
financial plans of the NIDDK and the NHLBI, the award of grants pursuant to this
PA is contingent upon the availability of funds for this purpose.

RESEARCH OBJECTIVES

Background

Mutations in the globin gene cluster are among the most common inherited diseases
in humans, leading to disorders that affect many individuals in the U.S. and
around the world, such as sickle cell anemia and thalassemia (Cooley's anemia). 
Treatment for these disorders eventually may include both pharmaceutical agents
and gene therapy. Some therapeutic approaches currently being studied are based
on the fact that a number of naturally occurring mutations, such as HPFH and
delta beta-thalassemia, are associated with significantly elevated levels of
fetal hemoglobin (Hb F) in adult RBC. This increase in Hb F can be
therapeutically beneficial, and modifies the clinical severity of
hemoglobinopathies associated with abnormal or deficient beta globin chains.  A
number of experimental drugs have been used successfully in some patients to
increase the level of fetal hemoglobin in adults and some children.  These
studies have established the feasibility of this therapeutic approach to diseases
with abnormal beta-globin production including sickle cell anemia and beta-
thalassemia.  However, not all patients respond to these treatments, their
applicability to other disorders is not established, and the molecular and
cellular basis for their mode of action remains unclear.  Hence further advances
are needed to broaden the range of possible therapeutic strategies for disorders
of globin synthesis. The elucidation of precise molecular mechanisms responsible
for increased Hb F production in these disorders could lead to the development
of novel future therapeutic approaches to treat hemoglobinopathies.  In addition,
studies of the developmental- and tissue-specific control of the globin gene
cluster are important for optimizing gene transfer, via viral or chemical
vectors, for gene replacement therapy.

In the last decade, the DNA of the human alpha- and beta-globin gene clusters
have been fully mapped and sequenced.  Similar work has been done for a number
of other species that serve as model systems.  The basic control of globin gene
expression at the transcriptional level, as controlled by proximal promoters, and
at post-transcriptional levels has partially been elucidated.  In addition, a
number of important trans-acting factors and the cis-acting DNA sequences with
which they interact have been described. These include the GATA family, EKLF, and
other DNA-binding proteins.  Studies using transfection assays and transgenic and
knockout methods have partially clarified mechanisms of transcriptional control,
including reciprocal gamma- and beta-globin gene interactions.  Other mechanisms
including RNA splicing processes, mRNA stabilizing elements and other post-
transcriptional mechanisms also have been studied.

Recent studies have shown that the locus control region (LCR) is a dominant
regulator of many aspects of globin gene expression.  LCRs are key genetic
regulators that place large sub-elements of chromosomes under a common control,
and hence allow chromosomal regions with distinct functions to be differentially
regulated.  There is a fundamental need to clarify fully the elements involved
in the regulation of the expression of globin genes through its LCR.  In addition
to the benefits of this knowledge to disorders of hemoglobin production, this
system may serve as a model for advances in understanding the coordinate
regulation of other chromosomal gene clusters, and to answer fundamental
questions about chromatin structure and regulation of gene expression. Research
into the globin LCR is anticipated to expand our understanding of large-scale
chromosomal regulatory mechanisms operational over much of the genome.  The
nature and extent of interactions between the cis-acting sequences in the
proximal promoter and their trans-acting factors of the individual globin genes
and distal elements including the LCR remain controversial.  Mechanisms for
interaction or communication between distal elements and these promoters have not
been established definitively, nor has the mechanism of interaction of these
control regions with the transcription initiation complex been studied in any
detail.  The way in which cell cycle control, erythroid cell differentiation and
pharmacological agents, such as hydroxyurea, affect globin gene expressions also
are poorly understood.

 In addition to its fundamental importance, studies in this area are anticipated
to contribute to design of approaches to expand the erythroid compartment in bone
marrow or to expand erythroid precursors in vitro and to promote their
appropriate differentiation. The linkage between erythroid cell differentiation
and globin gene control needs further study.  The temporal processes dictating
globin gene expression and the mechanisms for the developmental timetable are
important unresolved issues in erythropoiesis of relevance to this initiative.

Research Goals and Scope

The purpose of this program announcement is to stimulate new avenues of research
into the developmental processes involved in the differential expression of
globin genes.  Applications from new investigators and investigators with
relevant expertise in other fields are encouraged.

The areas of research within this program announcement include, but are not
restricted to:

(1) Determine the chromatin structure associated with the active and repressed
(or silenced) states of the globin genes;

(2) Study the interaction of the LCR and other distal elements with the promoter
and transcription initiation complex of the individual globin genes;

(3) Determine the enzymatic activities needed for the structural transitions of
the chromatin associated with the active and repressed states of the globin
genes, and the proteins needed to maintain those states;

(4) Identify and characterize trans-acting factors and other proteins involved
in the developmental regulation of the globin genes;

(5) Characterize post-transcriptional mechanisms that may control developmental
control of globin genes;

(6) Examine the relevance of cell cycle control in erythroid differentiation and
globin gene expression;

(7) Study molecular and cellular mechanisms, operative in various high HbF
syndromes, that can modulate the production of HbF in all cells or increase the
number of circulating red cells that contain HbF (so-called F-cells);

(8) Determine the mechanism of action of drugs, such as hydroxyurea and butyrate,
that affect HbF levels;

(9) Develop new technological approaches to examine the mechanism of globin gene
regulation, especially with regard to the LCR, and new in vivo and in vitro
assays;

(10)  Identify non-globin-linked genes that may affect the expression of the
gamma gene;

(11)  Study the linkage between erythroid cell differentiation and the
developmental control of the globin genes;

(12)  Develop new model systems to study globin gene regulation.

In applying for support under this announcement, investigators are encouraged to
present other research ideas not suggested above.

INCLUSION OF WOMEN AND MINORITIES IN RESEARCH INVOLVING HUMAN SUBJECTS

It is the policy of the NIH that women and members of minority groups and their
sub-populations must be included in all NIH supported biomedical and behavioral
research projects involving human subjects, unless a clear and compelling
rationale and justification is provided that inclusion is inappropriate with
respect to the health of the subjects or the purpose of the research.  This
policy results from the NIH Revitalization Act of 1993 (Section 492B of Public
Law 103-43).

All investigators proposing research involving human subjects should read the
"NIH Guidelines For Inclusion of Women and Minorities as Subjects in Clinical
Research," which have been published in the Federal Register of March 28, 1994
(FR 59 14508-14513) and in the NIH Guide for Grants and Contracts, Volume 23,
Number 11, March 18, 1994.

Investigators may obtain copies of the policy from the program staff listed under
INQUIRIES.  Program staff also may provide additional relevant information
concerning the policy.

NIH POLICY AND GUIDELINES ON THE INCLUSION OF CHILDREN AS PARTICIPANTS IN
RESEARCH INVOLVING HUMAN SUBJECTS

It is the policy of NIH that children (i.e., individuals under the age of 21)
must be included in all human subjects research, conducted or supported by the
NIH, unless there are scientific and ethical reasons not to include them.  This
policy applies to all initial (Type 1) applications submitted for receipt
dates after October 1, 1998.

All investigators proposing research involving human subjects should read the
"NIH Policy and Guidelines on the Inclusion of Children as Participants in
Research Involving Human Subjects" that was published in the NIH Guide for Grants
and Contracts, March 6, 1998, and is available at the following URL address:
http://www.nih.gov/grants/guide/notice-files/not98-024.html

APPLICATION PROCEDURES

Applications are to be submitted on the grant application form PHS 398 (rev.
5/95) and will be accepted at the standard application deadlines as indicated in
the application kit.  Application kits are available at most institutional
offices of sponsored research and may be obtained from the Division of Extramural
Outreach and Information Resources, National Institutes of Health, 6701 Rockledge
Drive, MSC 7910, Bethesda, MD 20892-7910, telephone 301-435-0714, email:
asknih@od.nih.gov.

The program announcement title and number must be typed on line 2 of the face
page of the application form and the YES box must be marked.

The completed original application and five legible copies must be sent or
delivered to:

CENTER FOR SCIENTIFIC REVIEW
NATIONAL INSTITUTES OF HEALTH
6701 ROCKLEDGE DRIVE, ROOM 1040 - MSC 7710
BETHESDA, MD  20892-7710
BETHESDA, MD  20817 (for express/courier service)

REVIEW CONSIDERATIONS

Applications will be assigned on the basis of established Public Health Service
referral guidelines.  Applications that are complete will be evaluated for
scientific and technical merit by an appropriate peer review group convened in
accordance with NIH peer review procedures.  As part of the initial merit review,
all applications will receive a written critique and undergo a process in which
only those applications deemed to have the highest scientific merit, generally
the top half of applications under review, will be discussed, assigned a priority
score, and receive a second level review by the appropriate national advisory
council or board.

Review Criteria

The goals of NIH-supported research are to advance our understanding of
biological systems, improve the control of disease, and enhance health.  The
reviewers will comment on the following aspects of the application in their
written critiques in order to judge the likelihood that the proposed research
will have a substantial impact on the pursuit of these goals.  Each of these
criteria will be addressed and considered by the reviewers in assigning the
overall score weighting them as appropriate for each application.  Note that the
application does not need to be strong in all categories to be judged likely to
have a major scientific impact and thus deserve a high priority score.  For
example, an investigator may propose to carry out important work that by its
nature is not innovative but is essential to move a field forward.

1.  Significance.  Does this study address an important problem? If the aims of
the application are achieved, how will scientific knowledge be advanced?  What
will be the effect of these studies on the concepts or methods that drive this
field?

2.  Approach.  Are the conceptual framework, design, methods, and analyses
adequately developed, well-integrated, and appropriate to the aims of the
project?  Does the applicant acknowledge potential problem areas and consider
alternative tactics?

3.  Innovation.  Does the project employ novel concepts, approaches or method? 
Are the aims original and innovative? Does the project challenge existing
paradigms or develop new methodologies or technologies?

4.  Investigator.  Is the investigator appropriately trained and well suited to
carry out this work?  Is the work proposed appropriate to the experience level
of the principal investigator and other researchers (if any)?

5.  Environment.  Does the scientific environment in which the work will be done
contribute to the probability of success?  Do the proposed experiments take
advantage of unique features of the scientific environment or employ useful
collaborative arrangements? Is there evidence of institutional support?

The initial review group also will examine: the appropriateness of proposed
project budget and duration; the adequacy of plans to include both genders,
minorities and their subgroups, and children as appropriate for the scientific
goals of the research and plans for the recruitment and retention of subjects;
the provisions for the protection of human and animal subjects; and the safety
of the research environment.

AWARD CRITERIA

Applications will compete for available funds with all other approved
applications.  The following will be considered in making funding decisions:

o  Quality of the proposed project as determined by peer review
o  Availability of funds
o  Program priority.

INQUIRIES

Inquiries concerning this PA are encouraged.  The opportunity to clarify any
issues or questions from potential applicants is welcome.

Inquiries regarding programmatic issues may be directed to:

David G. Badman, Ph.D.
Division of Kidney, Urologic and Hematologic Diseases
National Institute of Diabetes and Digestive and Kidney Diseases
45 Center Drive, Room 6AS-13C MSC 6600
Bethesda, MD  20892-6600
Telephone:  (301) 594-7717
FAX:  (301) 480-3510
Email:  David_Badman@nih.gov

Helena O. Mishoe, Ph.D.
Division of Blood Diseases and Resources
National Heart, Lung and Blood Institute
6701 Rockledge Drive, Room 10156
Bethesda, MD  20892-7950
Telephone: (301) 435-0050
FAX:  (301) 480-0868
Email:  hm31y@nih.gov

Inquiries regarding fiscal and administrative matters may be directed to:

Aretina D. Perry-Jones
Division of Extramural Activities
National Institute of Diabetes and Digestive and Kidney Diseases
45 Center Drive, Room 6AN-38 - MSC 6600
Bethesda, MD  20892-6600
Telephone:  (301) 594-8862
FAX:  (301) 480-3504
Email:  PerryA@extra.niddk.nih.gov

Ms. Jane R. Davis
Grants Management Office
National Heart, Lung and Blood Institute
6701 Rockledge Drive, MSC 7926
Bethesda, MD  20892-7926
Telephone:  (301) 435-0166
FAX:  (301) 480-3310
Email:  jane_davis@nih.gov

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic Assistance Nos.
93.849 and 93.839.  Awards will be made under authority of the Public Health
Service Act, Title IV, Part A (Public Law 78-410, as amended by Public Law 99-
158, 42 USC 241 and 285) and administered under PHS grants policies and Federal
Regulations 42 CFR 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.

The PHS strongly encourages all grant and contract recipients to provide a smoke-
free workplace and promote the non-use of all tobacco products.  In addition,
Public Law 103-227, the Pro-Children Act of 1994, prohibits smoking in certain
facilities (or in some cases, any portion of a facility) in which regular or
routine education, library, day care, health care or early childhood development
services are provided to children.  This is consistent with the PHS mission to
protect and advance the physical and mental health of the American people.


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