Subject: RFA HL-9l-05-H Full Text
From: Dot Baker <[email-masked]>
To: [email-masked]
Date: Tue, 02 Apr 91 13:43 EST

Hi Paul,
    Another one.
                Dottie
 ---------------------------- Text of forwarded message -----------------------
 
$$XID RFA HL9105H HL-91-05-H P1O1 **************************************
 
REQUEST FOR APPLICATIONS
 
RFA:  HL-91-05-H
 
P.T. 34; K.W. 0715115, 0780015, 1002004, 1002008
 
CELL LINES FOR HYPERTENSION RESEARCH
 
National Heart, Lung, and Blood Institute
 
Letter of Intent Receipt Date:  August 2, 1991
Application Receipt Date:  September 6, 1991
 
PURPOSE
 
The objective of this Request for Applications (RFA) is to
encourage research to develop and to characterize primary or
immortalized cell lines applicable to the study of hypertension.
Particular emphasis is placed on maintenance of the phenotype of
cells derived from adrenal, endothelial, renal, and vascular
tissues.  Specific phenotypes include hormone production and/or
responsiveness, transport functions, expression of key structural
genes, or other properties relevant to the study of blood
pressure regulation.
 
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 Lines for Hypertension Research," is
related to the priority area of Heart Disease and Stroke.
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, D.C. 20402-9325 (telephone 202-783-3238).
 
BACKGROUND
 
Cell culture is an essential tool of modern biological research
that is particularly useful for the investigation of physiologic
and pathologic processes and mechanisms associated with human
diseases.  Hypertension is a multifactorial aberration of a
complex interaction among numerous physiological systems and
regulatory cascades.  In vitro systems, such as cultured cells,
provide a way to dissect the precise role of individual cellular
components.  Application of cell culture technologies to the
study of cardiovascular health and disease has been hindered both
by the limited types of clonal cell lines currently available and
dedifferentiation of primary cells maintained in culture.  These
drawbacks prevent the use of cultured cells in many instances in
which a particular phenotype found in vivo is to be studied.
 
The development of techniques that maintain or promote cell
differentiation would be of great value in many areas of blood
pressure research because it would allow better control of the
environment of the experimental preparation, the use of
homogeneous cell populations with predictable characteristics for
long-term experiments, and the introduction of molecular
biological techniques allowing the expression of selected genes.
Moreover, the development of cell lines that stably maintain
desired phenotypes would also decrease the need for intact animal
models and, therefore, reduce the use of laboratory animals.
 
Cells isolated from adrenal, endothelial, renal, and vascular
tissues have provided models for the study of biological
mechanisms involved in the pathophysiology of hypertension.
Unfortunately, primary cell cultures invariably contain multiple
cell types and frequently lose several phenotypes present in
vivo.  For instance, vascular smooth muscle cells rapidly lose
their contractile phenotype.  This function can only be partially
recovered under appropriate culture conditions.  Endothelial
cells lose responsiveness to acetylcholine that stimulates the
generation of endothelium derived relaxing factor.  Similarly,
cultured renal epithelial cells isolated from the rabbit thick
ascending limb of Henle do not retain the full expression of
cytochrome P450 activity found in freshly isolated tissues, nor
do they express fully the transport activities that are the
important differentiated characteristics.  Further,
juxtaglomerular cells are difficult to isolate.  Once isolated
and cultured, they lose their ability to process and secrete
renin.
 
Culture conditions:
 
The development, in the mid-1970s, of serum-free, chemically
defined media for cultured cells provided a new impetus for
research utilizing cell cultures.  In the absence of serum, it
becomes possible to define the nature of various growth factors
and to identify factors responsible for maintenance or induction
of differentiation.  In fact, serum factors inhibit induction of
cellular differentiation and enhance oncogene-induced
transformation.  Several recent studies show that it is possible
to induce or maintain differentiated characteristics in cultured
cells by controlling the cell density, adding appropriate
hormonal agents, avoiding the use of proteases during cell
isolation, employing culture conditions that improve nutrient and
gas exchange and providing appropriate extracellular matrices.
However, the aforementioned studies have only met with partial
success and have not fully defined the factors that promote cell
differentiation.
 
Molecular approaches:
 
Immortalization of well characterized primary cell cultures can
be accomplished by infection with oncogenic viruses or by
transfection with appropriate vectors.  However, studies suggest
that these techniques require a mitotic target cell.  For
example, infection and immortalization of primary cells with
retroviruses requires that the host cell undergo an S phase;
post-mitotic cells generally do not integrate or yield stable
expression of these transforming agents.  Immortalization and/or
transformation of cells by oncogenes is often associated with
loss of differentiated characteristics.  Immortalization of
progenitor cells followed by arrest of oncogene expression and
application of conditions that promote differentiation of desired
phenotypes have circumvented some of these difficulties.  For
example, immortalized murine neural precursor cells have been
developed that differentiate into astrocytes by stimulation with
appropriate factors.  Mitotic cells immortalized with temperature
sensitive alleles of several oncogenes have further permitted
cell growth at permissive temperatures and differentiation
induced at non-permissive temperatures.  Application of these
technologies to post-mitotic cells such as adrenal, endothelial,
renal and vascular cells should improve the potential for
development of immortalized lines.  The effectiveness of
different DNA constructs and various DNA transfection methods in
generating immortalized cells needs to be explored and evaluated
for efficacy in each particular cell type.
 
Recent developments indicate that it may be possible to use
transgenic animals to obtain immortalized cell lines with
appropriate characteristics.  Fusion genes are constructed so
that expression of an oncogene is targeted to a particular cell
type.  Cell lines can then be derived from tumors that result
from the expression of the oncogene.  Major progress in
understanding the genetic and molecular basis of cell growth and
hypertrophy comes from the isolation and characterization of
cellular oncogenes.  Several oncogenes (c-myc, SV40 large T
antigen, c-Ha-ras, v-Ha-ras) have been microinjected into the
pronuclei of fertilized mouse eggs to produce transgenic mice
bearing active oncogenes that are stably integrated into their
genome.  These animals develop malignancies in many diverse
tissues at high frequencies.  Construction of tissue-specific
promoters fused to oncogenes and introduction of these into the
germline using a transgenic approach permits targeting of
activated oncogenes to particular tissues and to particular cells
within a tissue.  For example, rat elastase I regulating elements
directed c-Ha-ras to the exocrine pancreas, leading to neoplastic
transformation of differentiating acinar cells, while
insulin-promoted SV 40 large-T-antigen yielded beta-cell tumors.
These experiments suggest that oncogenes targeted to particular
cells via tissue-specific regulatory elements may permit
development of desired cell lines.
 
The new cell culture techniques and recent advances in molecular
biology described above can now be systematically applied to
establish cell lines suitable for hypertension research.
Development of transformed cell lines derived from adrenal,
endothelial, renal, and vascular tissues will only be useful if
lines continue to display phenotypes typical of the cells from
which they are derived.  Identification of markers that have a
significant role in normal or aberrant regulation of blood
pressure must be demonstrated.  Cell lines currently available,
such as adrenal Y1 cells, are insufficient for many studies
because they lack hormone responsiveness and do not synthesize
aldosterone.  Similarly, LLCPK-1 cells, a porcine kidney cell
line that contains vasopressin receptors, lacks co-transport
functions.  While MDCK cells, a dog kidney cell line, remain
responsive to many hormones and retain their capability in fluid
transport, these cells no longer resemble any particular single
cell type from the kidney.  Development of cell lines capable of
producing renin, aldosterone, mitogens, and other vasoactive
agents, active in ion transport, and responsive to various
relevant hormones and other blood-borne substances, would
facilitate basic studies on the function of these cells in
physiologic and pathologic regulation of blood pressure.
 
OBJECTIVES AND SCOPE
 
The proposed studies would place emphasis on adrenal,
endothelial, renal, and vascular cells.  Examples of appropriate
research areas are given below.  However, the list is not meant
to be complete or to exclude other relevant research.  The goals
to be addressed are:
 
1) Development of culture conditions that maintain or induce the
expression of selected biological characteristics in cells
relevant to hypertension research.
 
2) Development of immortalized cell lines having appropriate
phenotypes, including, but not limited to, transport functions,
production of vasotropic and vasoactive substances, and response
to these hormones.
 
3) Introduction and expression of specific genes, relevant to the
study of hypertension, in cultured cells in a regulatable
fashion.
 
4) Co-culture of well-defined cell lines to study cell-cell
communication and paracrine mechanisms responsible for the
regulation of blood pressure.
 
Cell lines must be developed specifically for the use of
understanding biological phenomena of importance to normal and
altered regulation of blood pressure.  Consequently, research
applications in response to this initiative must include specific
aims for understanding mechanisms relevant to blood pressure
homeostasis.
 
SPECIAL INSTRUCTIONS TO APPLICANTS REGARDING IMPLEMENTATION OF
NIH POLICIES CONCERNING INCLUSION OF WOMEN AND MINORITIES IN
CLINICAL RESEARCH STUDY POPULATIONS
 
NIH and ADAMHA policy is that applicants for NIH/ADAMHA 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 Section 2, A-D of the Research
Plan AND summarized in Section 2, E, Human Subjects.
Applicants/offerors are urged to assess carefully the feasibility
of including the broadest possible representation of minority
groups.  However, 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,
prevention (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.
 
EXCLUSIONS
 
Although multidisciplinary approaches are encouraged, it is not
the intent of this announcement to solicit applications for large
studies that would encompass a variety of independent projects,
i.e., program projects.  This program will not support clinical
trials or large epidemiological studies.  In general, funds will
not be provided for the purchase and installation of expensive,
new equipment.  Awards under this announcement to foreign
institutions will be made only for research of very unusual
merit, need and promise, and in accordance with Public Health
Service policy governing such awards.
 
MECHANISM OF SUPPORT
 
The support mechanism for this program will be the traditional,
individual research grant (RO1).  Although approximately $1.25
million in total costs for this program is included in the
financial plans for fiscal year 1992, award of grants pursuant to
this RFA is contingent upon receipt of funds for this purpose.
It is anticipated that five (5) grants will be awarded under this
program.  The specific number to be funded, however, will depend
on the merit and scope of the applications received and 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.
 
Applications may be submitted by for-profit or non-profit
organizations, whether public or private, such as universities,
colleges, hospitals, laboratories, units of State or local
governments, and eligible agencies of the Federal government.
Applications from minority individuals and women are encouraged.
 
Upon initiation of the program, the Division of Heart and
Vascular Diseases will sponsor periodic meetings to encourage
exchange of information among investigators who participate in
this program and to stimulate collaboration.  Applicants should
request ADDITIONAL TRAVEL FUNDS for a one-day meeting each year,
most likely to be held in Bethesda, Maryland.  Applicants should
also include a statement in their applications indicating their
willingness to participate in these meetings.
 
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 National Institutes of Health (NIH).
It is anticipated that support for the present program will begin
in May 1992.  Administrative adjustments in project period and/or
amount of support may be required at the time of the award.
 
If collaborative arrangements involve sub-contracts with other
institutions, the NHLBI Grants Operation Branch should be
consulted regarding procedures to be followed (Tel:
301-496-7257).
 
All current policies and requirements that govern the research
grant programs of the NIH will apply to grants awarded under this
RFA.
 
REVIEW PROCEDURES AND CRITERIA
 
Review Method:  Upon receipt, applications will be reviewed for
their responsiveness to the objectives of this RFA.  If an
application is judged unresponsive, the applicant will be
contacted and given the opportunity to withdraw the application,
or have it considered for the regular, investigator-initiated
grant program of the NIH.  If the application submitted in
response to this RFA is substantially similar to a research 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.
 
Applications judged to be responsive will be reviewed for
scientific and technical merit by an initial review group
convened by the Division of Extramural Affairs, NHLBI, solely to
review these applications.
 
This initial review will include a preliminary evaluation to
determine scientific merit relative to the other applications
received in response to this RFA (triage).  The NIH will withdraw
from further consideration applications judged to be
noncompetitive and promptly notify the Principal
Investigator/program director and the official signing for the
applicant organization.  Those applications judged to be
competitive will be further evaluated for scientific/technical
merit by the usual peer review procedures.
 
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 traditional research project grant
applications, including:
 
o the novelty, originality, and feasibility of the approach and
the adequacy of the experimental design, including the
suitability and composition of the study population
 
o the competence of the Principal Investigator and collaborators
to accomplish the proposed research and the commitment and time
they will devote to the project
 
o the suitability of the facilities to perform the proposed
research, including laboratories, instrumentation, and data
management systems
 
o the appropriateness of the requested budget for the work
proposed
 
o adequate plans for interaction and communication of information
and concepts among investigators involved in collaborative
studies
 
METHOD OF APPLICATION
 
Letter of Intent:  Prospective applicants are asked to submit a
letter of intent to apply to this RFA.  This letter should
include the names of any participating institutions and all
investigators, together with a descriptive title.  Such a letter
of intent is not binding, it will not enter into the review of
any application subsequently submitted, nor is it a necessary
requirement for application.  Letters of intent are requested
solely for planning purposes.  The NHLBI Staff will not provide
responses to such letters.  Prospective applicants are asked to
submit a letter of intent to apply to this RFA by August 2, 1991,
and be addressed to:
 
Dr. Charles L. Turbyfill
Review Branch, Division of Extramural Affairs
National Heart, Lung, and Blood Institute
Westwood Building, Room 553-A
National Institutes of Health
Bethesda, MD  20892
 
Format for Applications:  Submit applications on form PHS 398
(revised 10/88), the application form for the traditional NIH
research project grant.  Copies of this form are available in the
applicant institution's office of sponsored research and may be
obtained from the following:
 
Office of Grants Inquiries
Division of Research Grants
Westwood Building, Room 449
National Institutes of Health
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
above on "Review Procedures and Criteria" are fulfilled.  To
identify the application as a response to this RFA, CHECK "YES"
on item 2 of page 1 of the application and enter the title "Cell
Lines for Hypertension Research" and enter the RFA number
HL-91-05-H in the space provided.
 
THE RFA LABEL FOUND IN THE PHS 398 APPLICATION KIT MUST BE
AFFIXED TO THE BOTTOM OF THE FACE PAGE OF THE ORIGINAL COMPLETED
APPLICATION.  FAILURE TO USE THIS LABEL COULD RESULT IN DELAYED
PROCESSING OF YOUR APPLICATION SUCH THAT IT MAY NOT REACH THE
REVIEW COMMITTEE IN TIME FOR REVIEW.
 
Application Procedure:  Send or deliver the completed, signed
application and four (4) complete photocopies to the following,
making sure that the original application with the RFA label
attached is on top:
 
Division of Research Grants
Westwood Building, Room 240
National Institutes of Health
Bethesda, MD  20892**
 
SEND TWO ADDITIONAL COPIES OF THE 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 FOUR 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 September 6, 1991.
 
An application not received by this date will be  considered
ineligible and will be returned to the applicant.
 
Timetable:
 
Letter of Intent ........................... August 2,  1991
Application Receipt Date ................  September 6, 1991
Review by National Heart, Lung, and
     Blood Advisory Council ............. February 6-7, 1992
Anticipated Award Date ......................... May 1, 1992
 
Inquiries:  Inquiries regarding this announcement may be
directed to the following:
 
Dr. Michael C. Lin
Hypertension and Kidney Diseases Branch
Division of Heart and Vascular Diseases
National Heart, Lung, and Blood Institute
Federal Building, Room 4C10
National Institutes of Health
Bethesda, MD  20892
Telephone:  (301) 496-1857
Fax:  (301) 496-9882
 
For fiscal and administrative matters, contact:
 
Jane R. Davis
Grants Operations Branch
Division of Extramural Affairs
National Heart, Lung, and Blood Institute
Westwood Building, Room 4A15C
Bethesda, MD  20892
Telephone:  (301) 496-7255
 
This program is described in the Catalog of Federal Domestic
Assistance number 93.837, Heart and Vascular Diseases.  Awards
will be made under the authority of the Public Health Service
Act, Section 301 (42 USC 241) and administered under PHS grant
policies and Federal regulations, most specifically 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 review by a Health Systems Agency.