From: kristoff@GENBANK.BIO.NET (Dave Kristofferson)
Newsgroups: bionet.sci-resources
Subject: NIH Guide, vol. 21, no. 32, pt. 2, 4 September 1992
Date: 15 Sep 92 18:54:17 GMT
Distribution: bionet


$$XID RFA CA9214 CA-92-14 P1O1 *****************************************

BIOMARKERS OF DIETARY FAT IN POST-MENOPAUSAL WOMEN

NIH GUIDE, Volume 21, Number 32, September 4, 1992

RFA:  CA-92-14

P.T. 34; K.W. 0710095, 0760003, 0765025

National Cancer Institute

Letter of Intent Receipt Date:  October 7, 1992
Application Receipt Date:  January 26, 1993

PURPOSE

The Division of Cancer Prevention and Control (DCPC), National Cancer
Institute (NCI), invites applications for cooperative agreements
directed towards the identification and evaluation of potential
biochemical/biological markers to assess total dietary fat intake in
post-menopausal women and to monitor adherence to dietary interventions
in cancer prevention clinical trials.

A major challenge in studying the effects of diet on health and chronic
disease risks is the difficulty of assessing dietary intake.  Current
methods for dietary assessment and adherence monitoring have different
levels of precision and accuracy and all have the inherent limitation
of relying on self-reported data.  The availability of
biochemical/biological markers for assessing dietary intake and
adherence monitoring would greatly facilitate the design, conduct, and
interpretation of cancer prevention clinical trials, analytical
epidemiologic studies and diet-health survey studies on the
relationship of diet and cancer risk.

The specific objectives of this Request for Applications (RFA) are to
encourage research on the identification and evaluation of
biochemical/biological indicators of total fat intake in
post-menopausal women on self-selected low- and high-fat diets and in
controlled feeding and/or metabolic studies.

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,
Cooperative Agreement for Biomarkers of Dietary Fat in Post-Menopausal
Women, is related to the priority area of cancer prevention.  Potential
applicants may obtain a copy of "Healthy People 2000" (Full Report:
Stock No. 017-001-00474-0 or Summary Report:  Stock No. 017-001-0043-1)
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 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.
Applications from minority individuals and women are encouraged.

MECHANISM OF SUPPORT

This RFA will support awards through the cooperative agreement (U01)
mechanism.  An assistance relationship will exist between the NCI and
the awardees to accomplish the purpose of the activity.  The planning,
direction, and execution of the proposed research will be the
responsibility of the awardees, with involvement of the NCI as
described in SPECIAL REQURIREMENTS.

In addition to the requirements stated in this RFA, awards will be
administered under PHS grants policy as stated in Public Health Service
Grants Policy Statement, DHHS Publication No. (OASH) 90- 50,000,
revised October 1, 1990.

This RFA is a one-time solicitation.  Future unsolicited competing
continuation applications will compete with all investigator-initiated
applications and be reviewed by the Division of Research Grants (DRG).
However, should the NCI determine that there is a sufficient continuing
program need, the NCI may invite all funded recipients to submit
competing continuation applications.

FUNDS AVAILABLE

Approximately $750,000 in total costs per year for three years will be
committed to fund applications that are submitted in response to this
RFA.  It is anticipated that three to five awards will be made.  This
level of support is dependent on the receipt of a sufficient number of
applications of high scientific merit.  The total project period for
applications submitted in response to the present RFA may not exceed
three years.

RESEARCH OBJECTIVES

The purpose of this RFA is to encourage the submission of applications
from qualified investigators interested in conducting investigations
designed to identify, characterize, and evaluate biochemical/biological
markers to assess dietary fat intake and adherence in post-menopausal
women.  This research initiative also seeks to promote the interactions
of investigators with scientific expertise, facilities, and
capabilities to conduct controlled feeding studies, metabolic studies,
and field studies.

Background

Present knowledge of the relationship of diet to human health and
chronic diseases has been derived from observational epidemiologic
studies.  There are marked variations in the international incidence
and mortality rates of various cancers and in the per capita
availability of fat, and correlations between these suggest that
dietary intake is an important risk factor.  Support for an etiologic
relationship between dietary fat and breast cancer is provided by
migrant population studies and by experimental animal studies which
have shown that high fat diets enhance mammary tumor development and
growth.

The results of analytical epidemiologic studies (case-control and
cohort) on the association of dietary fat with cancer incidence have
been inconsistent.  Some studies have shown a strong association of
dietary fat with the incidence of some cancers whereas in others there
is evidence of either a weak or no association.  In the epidemiologic
studies on dietary fat and cancer that have shown a positive
association, the results generally indicate that higher intakes of
total fat as well as saturated fat are associated with higher incidence
and mortality rates of certain cancers.  Inconsistent findings such as
these are difficult to interpret.  Measurement errors of dietary intake
in general and of fat intake in particular, may in part account for
these inconsistencies.

Methods commonly used in epidemiologic research to assess dietary
intake include per capita food availability, household food
inventories, and food records and dietary recalls, both of which
include diet histories and food frequency questionnaires.  Per capita
food availability provide useful leads for further research on the
relationship of diet to chronic diseases because they enable
investigators to compare rates of disease among countries with marked
differences in mortality and incidence rates and in the availability of
specific nutrients in their food supply.  However, these
cross-sectional comparisons do not control for confounding factors, nor
can they be used to show associations between diet and disease in
individuals.

The choice of the dietary assessment method depends on whether the
assessment pertains to the average intake of the group or to the usual
intakes of individuals within a group, the level of detail (e.g., food
groups, foods, or nutrients) desired, and the degree of precision
needed in determining amounts of foods and nutrients consumed.
Additional considerations include the costs, burden on respondents, and
availability of trained interviewers.

Diet history methods rely on interviewers or questionnaires to estimate
the usual diet or habitual intake.  Some diet histories are obtained
through questionnaires, administered by an interviewer or completed
independently by the respondent.  This is often referred to as the
foods frequency questionnaire in which the number of times each listed
food is consumed and sometimes the portion sizes or amounts are
reported.  Often, many foods consumed may not be listed on the food
frequency questionnaire.

The 24-hour recalled method requires that respondents report the types
and amounts of foods they consumed over the previous 24-hour period.
Information is obtained by face-to-face interview or by telephone
interview.

The food record method requires participants to measure and record
types and amounts of all foods and drinks consumed over a specified
number of days, usually three to seven.  In some studies, all foods are
weighed, while in others, measuring cups and spoons and a ruler are
used to assess dimensions.  Food models, volume models, and photographs
are sometimes used to estimate portion sizes.

Accuracy can be reduced by many factors, such as poor memory of past
dietary practices, inaccurate recall of foods eaten and the amounts,
and the desire to please an interviewer.  Therefore, true intake can
only be determined with certainty if actual intakes are observed or
measured and weighed.  This has been feasible in only a few studies of
small numbers of subjects conducted for a short time.  Since true
dietary intake can rarely be determined, investigators often try to
assess the accuracy of a new or modified method by comparing results
not with true intake but with results from some other accepted but
possibly inaccurate method.

Several types of errors may occur in the current state-of-the-art
methods to assess dietary intake.  These include sampling errors,
reporting errors, errors due to wide day-to-day variation in dietary
intake, interviewer bias, and errors due to use of food composition
tables employed in estimating nutrient intakes.

Assessing nutrient intake is also limited by the accuracy and
completeness of standard food composition tables.  There are
sufficiently accurate data on the composition of protein and fat in
most foods, but for dietary fiber, some vitamins, and trace values in
food composition tables, averages of representative samples of a given
food are used and do not indicate the nutrient content of any specific
food sample.

Two recent reports, "The Surgeon General's Report on Nutrition and
Health" and the National Research Council's report "Diet and Health"
have listed as specific research recommendations the need for
identification and development of biochemical markers of dietary intake
and of exposure to dietary fats.  Identification of
biochemical/biological indicators of dietary exposure would circumvent
the current dietary assessment methodological short-comings that limit
the interpretation of data and often prevent the derivation of precise
conclusions about the association of dietary patterns and/or specific
dietary components with the risk of cancer and other chronic diseases.

The fatty acid composition of plasma and body tissues including
erythrocytes, platelet, adipose tissue and cheek cells have been used
to assess the types of fatty acids consumed.  However, no single
parameter or battery of biochemical/biological parameters have yet been
identified to reflect total fat intake, to validate self-reported
dietary intake, or to monitor adherence to low-fat diets.

Specific and sensitive biochemical/biological indicators of dietary
intake, in particular total dietary fat, would greatly facilitate the
design, conduct, and interpretation of dietary intervention trials,
epidemiologic studies and diet-health survey studies that attempt to
determine the role of diet in cancer risk and prevention.  In studies
involving dietary modifications, the extent to which the results may be
influenced by varying degrees of adherence is an aspect that is both
important and difficult to evaluate.  Thus, evaluation and validation
of potential biochemical/biological indices of dietary intake will
require controlled human feeding studies using well-defined diets and
precise measures of actual intake.

Studies of Special Interest

Specifically, applications are solicited that will: (1) identify and
evaluate potential biochemical/biological indicators of adherence in
post-menopausal women on self-selected low- and high-fat diets, and/or
(2) identify and evaluate biochemical/biological indicators of
adherence to low-fat diets in post-menopausal women in controlled
feeding and/or metabolic studies.  Applicants may also propose to
conduct a series of short-term (6-12 weeks) controlled clinical and/or
metabolic studies and/or field studies.  These studies should be
designed to identify, characterize, and evaluate minimally invasive,
specific, and sensitive biochemical/biological indicators for assessing
total fat intake and/or for monitoring adherence to low-fat diets.
Emphasis should be focused on dietary patterns that are nutritionally
adequate and are characterized by reduced levels of total fat and
saturated fat, increased levels of complex carbohydrates and fiber, and
include a variety of foods typically present in the U.S. diet.  In
addition, the influence of varying the levels of fat intake while
keeping fiber intake constant, weight loss, and energy balance may be
taken into consideration in the study designs.

Characteristics of an ideal biomarker may include the following, as
well as others: (1) specimens for analyte measurements are readily
accessible and involve minimally invasive procedures; (2) analyte is
stable during prolonged storage; (3) analyte assays are precise,
accurate and inexpensive; (4) analyte is sensitive to fat quantity and
quality; (5) dose-response range is detectable; (6) minimal intra- and-
inter-individual variation with defined intake; (7) minimal diurnal
variation/seasonal variation; (8) minimal sensitivity to confounders;
(9) defined time course of response and, (10) practical and low-cost.

SPECIAL REQUIREMENTS

Terms of Cooperation

The cooperative agreement award mechanism requires establishment of a
partnership and an assistance relationship between the NCI and award
recipients.  The role of the NCI will be to provide technical
assistance and to assist in the coordination of certain activities of
the award recipients as described below.  The "partner role" of the NCI
will be to facilitate, not to direct, the research activities.  The
Terms of Cooperation and other special requirements described below
will be incorporated in the Notice of Grant Award and 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, and NIH Grants Administration Policy Statements and other
NCI administrative terms of award.

A working group comprised of all Principal Investigators (PI) and the
NCI Program Director will meet as soon as possible after awards are
made to discuss approaches, to consider ways in which specific
protocols might be improved, to facilitate the comparability of certain
aspects of the research projects where desirable and feasible, to
identify areas of cooperation, and to facilitate the exchange of
information and material. Subsequently, the working group will meet
annually to exchange information and to seek means of resolving
problems that may have arisen.  The cooperative approach allows for
interactions among successful applicants and is designed to optimize
the use of resources, biological specimens, and methodologies.

The working group members may agree to share information and results in
order to open avenues of cooperation between investigators and to keep
abreast of the latest developments.  However, each awardee institution
will retain custody and primary rights to the data developed under
these cooperative agreements.

Responsibilities of NCI Program Director

The role of the NCI Program Director, who is Chief of the Diet and
Cancer Branch, DCPC, is to assist and facilitate, but not to direct,
research activities.  During the period of award the NCI Program
Director will schedule, coordinate, and actively participate in the
working group meetings.  The NCI Program Director will serve as a
liaison between the PIs and the NCI through annual meetings of the
working group and provide appropriate assistance to the investigators,
either individually or collectively as deemed necessary.

Through telephone contacts, written correspondence, and meetings, the
NCI Program Director will provide appropriate advice, assistance, and
guidance in identifying areas of potential cooperation and will assist
in facilitating the exchange of information and material.  The NCI
Program Director will assist the awardees with coordination of research
activities such as the exchange of biological specimens, analytical
methodologies, information, and results.  The NCI Program Director will
encourage standardization of analytical methodologies and sharing of
data between investigators when this is in the mutual interest of all
parties.  Coordination activities may also include the NCI Program
Director providing information on other ongoing studies and on NCI
priorities.

The NCI Program Director will monitor progress, review and evaluate
technical performance, and will review and approve all requests for key
personnel changes as appropriate during the project period.  Key
personnel are the PI and others specifically identified in the Notice
of Grant Award.

Responsibilities of Awardees

Awardees are responsible for the planning, direction, and execution of
the research projects, and for the dissemination and publication of
results in a timely manner.  The successful applicants will be
responsible for obtaining approval by the Institutional Review Board
(IRB) on all protocols with human subject involvement.  Informed
consent forms must comply with HHS Regulations and NIH guidelines.

The awardees are expected to establish quality assurance/quality
control procedures that will be an integral part of all laboratory
activities and measurements, dietary data collection, and compliance to
the dietary interventions.  The awardees agree to conduct the study
according to the IRB approved protocols and consent forms.

The awardees are expected to cooperate with each other by making
available information and results on a continuing basis and, where
practical, sharing methodologies and materials.  Data acquisition and
analyses are the responsibilities of the awardees.

Awardees are expected to participate in annual meetings of the working
group to be held in Bethesda, MD.  Applicants are advised to include
travel expenses to attend working group meetings in the budget requests
and to include a statement about willingness to participate in such
activities in the applications.  Copies of publications and major
presentations are to be provided to the NCI Program Director.

Publication of research results by the awardees is encouraged.  The
awardees will acknowledge NCI support in publications and oral
presentations of work supported by this cooperative agreement.

Arbitration Procedures

An arbitration panel of external consultants will be created as needed
to resolve any irreconcilable differences of opinion between the NCI
Program Director and the awardee(s) related to scientific/programmatic
matters or implementation of a proposed operating policy.  The panel
will include one award recipient designee, one NCI designee, and a
third designee chosen by the other two members of the arbitration
panel.

Reporting Requirements

Annual progress reports are to include a summary table of data and
results for minority study participants and a summary table for all
study participants combined.  In addition to the required annual
progress reports, the awardees will provide additional information
requested by the NCI Program Director at the discretion of the affected
PI(s).

Other Required Applicant Information

Applicants responding to the RFA will be expected to propose new and
innovative approaches to this research area and are reminded to
document the availability of multidisciplinary scientific expertise,
appropriate facilities, and the ability to recruit volunteer human
subjects including minorities.  Specifically, the applicants should
provide the following:  (1) multidisciplinary scientific expertise; (2)
appropriate facilities to conduct either controlled human feeding
studies and/or metabolic studies or field studies; (3) access to
existing cohorts of post-menopausal women with different dietary
patterns, e.g., customary high- and low-fat diets or participants
previously enrolled in other dietary modification research studies or
diet programs; (4) ability to recruit and enroll volunteer
post-menopausal women into controlled feeding studies and/or field
studies, and (5) access to laboratory facilities with capabilities of
performing analytical assays in biological specimens and foods.

Applicants from institutions which 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.  In such a case, a letter of agreement from either the GCRC
program director or PI must be included with the application.

STUDY POPULATIONS

SPECIAL INSTRUCTION TO APPLICANTS REGARDING IMPLEMENTATION OF NIH
POLICIES CONCERNING INCLUSION OF MINORITIES IN CLINICAL RESEARCH

The following is a statement of NIH and ADAMHA policy regarding the
inclusion of women and minorities in study populations.  Applications
that are responsive to this RFA will, be definition, meet the
requirement for inclusion of women.  The inclusion of minorities must
be addressed in applications submitted responding to this RFA.

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
Sections 1-4, of the Research Plan AND 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, 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 tissue 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

Prospective applicants are asked to submit, by October 7, 1992, a
letter of intent that includes a descriptive title of the proposed
research, the name and address of the PI, the names of other key
personnel, the participating institutions, and the number and title of
the RFA in response to which the application may be submitted.

Although a letter of intent is not required, is not binding, and does
not enter into the review of subsequent applications, it is requested
in order to provide an indication of the number and scope of
applications to be reviewed.

The letter of intent is to be sent to:

Postal mail address:

Carolyn K. Clifford, Ph.D.
Diet and Cancer Branch
National Cancer Institute
Division of Cancer Prevention and Control
9000 Rockville Pike
Executive Plaza North, Room 212
Bethesda, MD  20892-6130

Overnight delivery address:

Carolyn K. Clifford, Ph.D.
Diet and Cancer Branch
National Cancer Institute
Division of Cancer Prevention and Control
6130 Executive Plaza
Executive Plaza North, Room 212
Rockville, MD  20852

APPLICATION PROCEDURES

The research grant application form PHS 398 (rev. 9/91) is to be used
in applying for these grants.  These forms are available at most
institutional business offices; from the Office of Grants Inquiries,
Division of Research Grants, National Institutes of Health, Room 449,
Westwood Building, 5333 Westbard Avenue, Bethesda, MD  20892; and from
the NCI Program Director named below.

The RFA label available in the application form PHS 398 must be affixed
to the bottom of the face page.  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. In addition, the title of the
application, "Biomarkers of Dietary Fat in Post-Menopausal Women", and
the RFA number, CA-92-14, must be typed on line 2a of the face page of
the application form.

Submit a signed, typewritten original of the application, including the
Checklist, and three signed, exact photocopies in one package to the
DRG at the address listed below.  The photocopies must be clear and
single-sided.

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

At the time of submission, two additional copies of the application
must be sent to:

Referral Officer
Division of Extramural Activities
National Cancer Institute
Westwood Building, Room 828
5333 Westbard Avenue
Bethesda, MD  20892

Applications must be received by January 26, 1993.  If an application
is received after that date, it will be returned to the applicant. The
Division of Research Grants (DRG) will not accept any application in
response to this announcement that is essentially the same as one
currently pending initial review, unless the applicant withdraws the
pending application.  The DRG will not accept any application that is
essentially the same as one already 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

Upon receipt, applications will be reviewed by the DRG for
completeness.  Incomplete applications will be returned to the
applicant without further consideration.  Evaluation for responsiveness
to the program requirements and criteria stated in the RFA is an NCI
program staff function.  Applications that are judged non-responsive
will be returned, but may be submitted as investigator-initiated
research grants.  Questions concerning the responsiveness of proposed
research to the RFA should be directed to the Program Director
indicated in the LETTER OF INTENT section.

If the number of applications is large compared to the number of awards
to be made, the NCI may conduct a preliminary scientific peer review
(triage) to eliminate those applications that are clearly not
competitive.  The NCI will remove from competition those applications
judged to be noncompetitive for award and notify the applicant and
institutional business official.  Those applications judged to be both
competitive and responsive will be further evaluated according to the
review criteria stated below for scientific and technical merit by an
appropriate peer review group convened by the Division of Extramural
Activities, NCI.  The second level of review by the National Cancer
Advisory Board considers the special needs of the NCI and the
priorities of National Cancer Program.

The following factors will be considered in evaluating the scientific
merit of each response to this RFA:

o  Scientific merit, feasibility, and originality of the proposed
research;

o  Appropriateness and adequacy of the experimental approach and
methodology proposed to conduct the research;

o  Qualifications and research experience of the PI and key personnel;

o  Demonstration of availability of and access to potential study
participants and/or study population(s);

o  Adequacy of plans for inclusion of minorities;

o  Adequacy of provisions for the protection of human subjects;

o  Scientific plans and timetable for implementing the proposed
research and plans for effective interaction with other award
recipients and with the NCI;

o  Availability of resources necessary to perform the research, and

o  Adequacy of the proposed budget and duration in relation to the
proposed research.

AWARD CRITERIA

The anticipated date of award is September 30, 1993.  Awards made
pursuant to this RFA are contingent upon the availability of funds for
this purpose and the scientific merit review.

INQUIRIES

Written and telephone inquiries concerning the objectives and scope of
this RFA, and inquiries about specific proposed research are encouraged
and should be directed to the Program Director listed below.  The
program staff welcome the opportunity to clarify any issues or
questions from potential applicants.

Direct inquiries regarding programmatic issues to:

Carolyn K. Clifford, Ph.D.
Chief and Program Director
Diet and Cancer Branch
Division of Cancer Prevention and Control
National Cancer Institute
9000 Rockville Pike
Executive Plaza North, Room 212
Bethesda, MD  20892-6130
Telephone:  (301) 496-8573
FAX:  (301) 402-0553

Direct inquiries regarding fiscal matters to:

Eileen Natoli
Team Leader, PC Team
Grants Administration Branch
National Cancer Institute
6120 Executive Boulevard
Executive Plaza South, Room 243
Rockville, MD  20852
Telephone:  (301) 496-7800, ext.56

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic Assistance
No. 93.399, Cancer Control.  Awards will be made under the authority of
the Public Health Services Act, Title IV, Section 301 (Public Law
78-410,;42 U.S.C. 241 and Section 412, as amended by Public Law 99-518,
42 U.S.C 258a-1); 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 12732 or Health Systems Agency review.


$$XID RFA CA9222 CA-92-22 P1O1 *****************************************

POSSIBLE ROLE OF METALLOTHIONEIN IN CARCINOGENESIS

NIH GUIDE, Volume 21, Number 32, September 4, 1992

RFA:  CA-92-22

P.T. 34; K.W. 0715035, 1007009, 1003018, 1002008, 0710030

National Cancer Institute

Letter of Intent Receipt Date:  October 2, 1992
Application Receipt Date:  December 8, 1992

PURPOSE

The Chemical and Physical Carcinogenesis Branch, Division of Cancer
Etiology (DCE), National Cancer Institute (NCI), in collaboration with
the Cancer Biology Branch, Division of Cancer Biology, Diagnosis, and
Centers (DCBDC), NCI, and the Grants and Contracts Operations Branch,
Division of Cancer Treatment (DCT), NCI, invites investigator-initiated
research grant applications to elucidate the possible role of
metallothionein (MT) in carcinogenesis.  New and experienced
investigators in relevant fields and disciplines may apply for funds to
pursue this research.

Historically, three major groups of scientists have contributed to the
knowledge of MT: (1) toxicologists and physiologists interested in its
role in heavy metal metabolism and detoxification; (2) protein chemists
intrigued by its unusual structural features, and (3) molecular
biologists interested in gene regulation and the use of MT promoter
sequence for genetic engineering experiments.  In order to better
define the possible role of MT in carcinogenesis, this RFA seeks to
encourage interactions among these major groups, other basic
scientists, oncologists, and clinicians in multidisciplinary research
projects.

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), Possible Role of Metallothionein in
Carcinogenesis, is related to the priority area of cancer.  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 202-783-3238.

ELIGIBILITY REQUIREMENTS

Applications may be submitted by domestic and foreign, for-profit and
non-profit, public or private organizations, such as universities,
colleges, hospitals, laboratories, units of State and local
governments, and eligible agencies of the Federal Government.
Applications from women and minority investigators are encouraged.

MECHANISM OF SUPPORT

Support of this program will be through the National Institutes of
Health (NIH) traditional research project grant (R01).  Applicants will
be responsible for the planning, direction, and execution of the
proposed project.  The total project period for applications submitted
in response to this RFA may not exceed four years.  Awards 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, 1990.

This RFA is a one-time solicitation.  Future unsolicited competitive
continuation applications will compete with all other
investigator-initiated research grant applications and be peer reviewed
by a chartered study section in the Division of Research Grants (DRG),
NIH.  However, should the NCI determine that there is a sufficient
continuing program need, a request for competitive continuation
applications will be announced.  Only recipients of awards under this
RFA will be eligible to apply.

FUNDS AVAILABLE

Total costs of $1,500,000 per year for four years will be committed to
fund applications that are submitted in response to this RFA.  It is
anticipated that 9 to 11 awards will be made.  This funding level is
dependent upon the receipt of a sufficient number of applications of
high scientific merit.  The earliest feasible start date for the
initial awards will be July 1, 1993.  Although this program is provided
for in the financial plans of the NCI, the award of grants pursuant to
this RFA is also contingent upon the availability of funds for this
purpose.

RESEARCH OBJECTIVES

Background

Metallothioneins are a family of low molecular weight, cysteine-rich,
heavy metal-binding proteins.  The mammalian forms have 61 amino acids,
including 20 cysteine residues, but no aromatic amino acids or
histidines (1).  The metal content of purified MT is highly variable
and depends on the organism, tissue, and history of heavy metal
exposure.  For example, MT isolated from human liver autopsy samples
contains almost exclusively zinc (Zn), whereas MT from kidney contains
substantial levels of cadmium (Cd) and copper. These differences
probably reflect both the natural heavy metal exposure of the organs
and the expression of different MT isoforms (2).

The synthesis of MT in animals is induced in a variety of ways,
including exposure to heavy metals, physiologic stress, and changes in
endocrine status and requires increased transcription of MT genes
(3,4).  Our knowledge of MT degradation, however, is somewhat limited.
In vivo experiments have suggested that the rate of MT turnover in the
cytosol fraction is influenced by the specific metal bound to the
polypeptide.  The disappearance of 35S-labeled MT from the cytosol
fraction reflects degradation of this protein by lysosomal proteases
since apometallothionein (without metal), Zn-MT, and Cd-MT were rapidly
degraded in lysosomal extracts (5).

To date, our understanding of the function of MT has been focused on
its role in metal transport, mineral nutrition, metal detoxification,
and detoxification of other chemicals (6-8). Other possible functions
suggested for MT include participation in embryonic development, cell
differentiation and proliferation, and carcinogenesis (9-11).  In an
attempt to better understand the possible role of MT in carcinogenesis
and to develop future research directions in this area, a workshop was
sponsored by the Chemical and Physical Carcinogenesis Branch on
November 18-19, 1991.  The approach of the workshop was to focus only
briefly on what is known and to concentrate on what is unknown,
regarding MT and the cancer process.

The notion that MT may play a role in carcinogenesis is formulated on
the basis of two basic properties of this protein.  First, MT genes are
universally inducible.  A variety of stress conditions, excess metals
notwithstanding, has been shown to elicit expression of MT genes, which
are otherwise expressed at a relatively basal level.  Such a
transcriptional response is typical of cellular defense and suggests a
close relation with normal or abnormal growth.  Second, MT is
ubiquitous and highly conserved in its amino acid sequence wherever
cysteine residues are found.  These occur in motifs where cysteine
pairs are separated by one or two non-cysteine amino acids (CXC, CXXC)
(12).

On a strictly molecular affinity level, Cd can displace Zn from
Zn-fingers in DNA binding proteins (e.g., oncogenes, proto-oncogenes,
tumor suppressor genes), thereby altering their activity or function
within the cell.  Disruption of the function of these proteins could
result in an alteration in gene expression and thus in the promotion of
cell growth and turnover (promotion).  The displacement of Zn from
these proteins could generate free radicals thereby contributing to Cd
carcinogenesis.  In this regard, MT could function to sequester Cd,
thereby not permitting accumulation of free Cd within the cell, and
thus limiting the displacement of Zn from these important proteins.
The interaction of Cd with DNA binding proteins requires further
investigation.  In addition, such in vitro molecular research should
provide important insight into whole cell and whole animal systems.

The carcinogenic effect of Cd appears to be tissue specific and may be
related to MT levels in the target organs.  For example, the ventral
prostate, a target site of Cd carcinogenesis in the rat, appears to be
deficient in MT, thus apparently making it susceptible to
carcinogenesis by Cd.  Also, certain cells within the rat testes, and
possibly the lung, appear to contain low levels of MT, and both of
these tissues are clearly target sites for Cd carcinogenesis.  Further,
the ability to induce MT may be equally important in the carcinogenic
process.  Tissues able to respond rapidly to insult by producing MT may
achieve protection from the carcinogenic effects (e.g., liver), whereas
a "non-responsive" tissue (e.g., ventral prostate) may be highly
susceptible to Cd carcinogenesis.  Clearly, tissue differences in
susceptibility to the carcinogenic effects of Cd exist (13,14).  An
investigation of the differences between these tissues should provide
further insight into (1) a role of MT in Cd carcinogenesis and (2) a
possible mechanism of Cd carcinogenesis.  In addition, a species
difference in susceptibility to Cd exists, particularly within the
lung, and species comparisons may also provide answers to these
questions.

Most mammalian tissues contain a basal level of MT that may vary with
age and type of tissue.  Various localization techniques have
demonstrated MT mainly in the cellular cytosol, but it is also present
in the nucleus of hepatocytes in early development of the human fetus
and in fetal and neonatal rat.  Moreover, MT has been demonstrated in
various types of human thyroid, testicular germ cell tumors, bladder
transitional cell carcinomas, and salivary gland tumors.  Both benign
and malignant tumors express MT in the nucleus and/or cytoplasm and the
staining for MT is more predominant in cisplatin- and
radiation-resistant tumors than in those sensitive to chemotherapy.
The functional role of MT in protection of tumor cell growth or in drug
resistance is poorly understood.  A number of factors may be involved
in the development of drug resistance in tumor cells and expression of
MT may be one of them.  The importance of MT in this process appears to
depend on the phenotype of the tumor and its cellular origin (15,16).

Research Goals and Scope

The objectives of this RFA are to encourage research designed to
elucidate the possible role of MT in carcinogenesis.  Specific topic
areas that might be supported by the RFA include, but are not limited
to:

o  Biological and toxicological roles of MT.  Studies such as metal
homeostasis, detoxification, transport, role in cell proliferation
during the perinatal period, and involvement of Zn as a second
messenger in signal transduction, as related to cellular normality;

o  Regulation of MT gene expression.  Studies of metal induction in
various tissues, during development, and organismal specificity in
transgenic and model systems and in normal versus transformed cells;

o  Role of MT in tumor cell pathobiology.  Studies to define the role
of MT in tumor cell progression and metastasis and the types and
staging of tumors that may or may not express excess MT.  Studies
directed at enhancing a rational basis for therapeutic intervention
with metallic anticancer drugs;

o  Role of MT in cancer chemotherapy. Studies on the role of MT in
tumor cell resistance to anticancer drugs, especially metal based
drugs.  Studies on the use of induction of MT in non-tumor tissue as an
adjunct to reduce toxicity for metallic chemotherapeutics.  Studies
involving mechanisms by which MT synthesis could be specifically
depressed in tumor cells to make them hypersusceptible to metallic
chemotherapeutics;

o  Susceptibility factors in metal carcinogenesis.  Studies assessing
MT gene expression in target tissues of metallic carcinogens in rodents
and molecular epidemiology of MT with special emphasis on target
tissues of metallic carcinogens in humans (e.g., prostate, lung), and

o  Molecular interaction of MT with ligands (metals and anticancer
drugs) including binding and exchange, and structural and dynamic
studies.

These research objectives cut across the traditional administrative
divisions of the NCI.  The Chemical and Physical Carcinogenesis Branch,
DCE, is the issuing program.  The Cancer Biology Branch, DCBDC, and the
Grants and Contracts Operations Branch, DCT, are co-sponsoring
programs.  A joint funding plan will be developed after the review is
completed.

In addition, the National Institute of Environmental Health Sciences
has an interest in the general topic of metallothionein, but not in the
specific emphasis of this RFA, which is the possible role of
metallothionein in carcinogenesis.

STUDY POPULATIONS

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 are 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 must 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
and compelling rationale must 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 must be addressed in developing a research design
and sample size appropriate for the scientific objectives of the study.
This information must be included in the form PHS 398 (rev. 9/91) 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, and Hispanics.

The rationale for studies on single minority population groups should
be provided.

For the purpose of this policy, clinical research is defined as 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 the 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

Each prospective applicant is asked to submit, by October 2, 1992, a
letter of intent that includes a descriptive title of the proposed
research; the name, address, telephone and FAX numbers of the Principal
Investigator; the names of other key personnel; the participating
institution(s); and the number and title of the RFA in response to
which the application may be submitted.

Although a letter of intent is not required, is not binding, and does
not enter into the review of a subsequent application, it contains
information that is helpful in planning for the review.  The letter of
intent allows NCI staff to estimate the potential review workload and
helps to avoid conflict of interest in the review.

The letter of intent is to be sent to:

Dr. Yung-Pin Liu
Program Director, Carcinogenesis Mechanisms
Chemical and Physical Carcinogenesis Branch
Division of Cancer Etiology
National Cancer Institute
Executive Plaza North, Suite 700
Bethesda, MD  20892
Telephone:  (301) 496-5471
FAX:  (301) 496-1040

APPLICATION PROCEDURES

The research grant application form PHS 398 (rev. 9/91) is to be used
in applying for these grants.  The application package is available at
most institutional business offices; from the Office of Grants
Inquiries, Division of Research Grants, National Institutes of Health,
Westwood Building, Room 449, Bethesda, Maryland 20892, telephone
301-496-7441; and from the NCI Program Director named above.

The RFA label available in the application form PHS 398 must be affixed
to the bottom of the face page of the application.  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.  In addition,
the RFA number and title must be typed on line 2a of the face page of
the application form and the YES box must be marked.

Submit a signed, typewritten original of the application, including the
Checklist, and three signed, exact photocopies in one package to the
DRG at the address below.  The photocopies must be clear and single-
sided.

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

At the time of submission, two additional copies of the application
must be sent to:

Referral Officer
Division of Extramural Activities
National Cancer Institute
Westwood Building, Room 838
Bethesda, MD  20892

If the applicant has an approved assurance covering the research
(Multiple Project Assurance of Compliance for human subjects or Animal
Welfare Assurance for animal subjects), the applicant should provide
with the application certification of Institutional Review Board (IRB)
approval if humans are involved, and verification of Institutional
Animal Care and Use Committee (IACUC) approval if animals are involved.
These reviews and approvals should occur prior to submission of the
application, and the certifications and verifications should be
submitted with the application.  If animals or humans will be subjects
of the research at performance sites other than the applicant
organization, the applicant must identify in the application the
assurance status of each participant.  In cases where submission delays
are unavoidable, follow the instructions on pages 12 and 13 of the PHS
398 application package.  Failure to submit required certifications and
verifications to the Scientific Review Administrator of the initial
review group within 60 days of receipt of the application could result
in deferral or rejection.

Applications must be received by December 8, 1992.  If an application
is received after that date, it will be returned to the applicant.  The
DRG will not accept any application in response to this announcement
that is essentially the same as one currently pending initial review,
unless the applicant withdraws the pending application.  The DRG will
not accept any application that is essentially the same as one already
reviewed.  This does not preclude the submission of a substantial
revision to an already reviewed application, but such an application
must include an introduction addressing the previous critique.

REVIEW CONSIDERATIONS

Review Procedure

Upon receipt, applications will be reviewed by the DRG for
completeness.  An incomplete application will be returned to the
applicant without further consideration. Evaluation for responsiveness
to the RFA is an NCI program staff function.  Applications will be
judged to determine how well they meet the goals and objectives of the
program as described in the RFA.  Applications judged non-responsive
will be returned, but may be submitted as investigator-initiated
research grant applications at the next unsolicited receipt date.
Questions concerning the relevance of proposed research to the RFA are
to be directed to the Program Director as named in INQUIRIES.

If the number of applications is large compared to the number of awards
to be made, the NCI may conduct a preliminary scientific peer review to
triage applications on the basis of relative competitiveness and to
eliminate those applications that are clearly not competitive.  The NCI
will withdraw from further competition those applications judged to be
noncompetitive and notify the applicant and institutional business
official.

Those applications judged to be both competitive and responsive will be
further evaluated according to the review criteria stated below for
scientific and technical merit by an appropriate peer review group
convened by the Division of Extramural Activities, NCI.  The second
level of review by the National Cancer Advisory Board will consider the
special needs of the NCI and the priorities of the National Cancer
Program.

Review Criteria

Applications responsive to this competitive solicitation will be
reviewed in accordance with the criteria stated below:

o  scientific, technical or medical significance, and originality of
the proposed research;

o  appropriateness and adequacy of the experimental design methodology
proposed to carry out the research;

o  qualifications and research experience of the Principal Investigator
and proposed staff and/or collaborators, particularly but not
exclusively in the area of the proposed research as well as the level
of commitment and time availability of each on the project;
o  documentation of the availability and adequacy of existing/proposed
facilities, equipment and resources for all laboratories involved in
the research, including adequate laboratory safety, biohazard and
animal welfare    practices and procedures as well as provisions for
the protection of human subjects; and

o  appropriateness of the requested budget and timetable for completion
relative to the proposed research.

The review group will critically examine the submitted budget and will
recommend an appropriate budget and period of support for each scored
application.

AWARD CRITERIA

Awards will be made primarily on the basis of scientific merit but
responsiveness to the RFA, overall program balance, and the
availability of resources are important considerations that will be
taken into account.

INQUIRIES

The Program Director welcomes the opportunity to clarify any scientific
or programmatic issues or questions from potential applicants.  Written
and telephone inquiries concerning the objectives and scope of this
RFA, and inquiries about specific proposed research are encouraged and
are to be directed to Dr. Liu at the address provided in LETTER OF
INTENT.

Written and telephone inquiries of a budgetary, administrative, and/or
policy nature are to be directed to:

Ms. Jean Cahill
Grants Management Team Leader
Grants Administration Branch
National Cancer Institute
Executive Plaza South, Suite 243
Bethesda, MD  20892
Telephone:  (301) 496-7800, ext. 47
FAX:  (301) 496-8601

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic Assistance
No. 93.393, Cancer Cause and Prevention Research. Awards are made under
the authorization 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.

References

Kagi, J.H.R. and Schaffer, A. Biochemistry of metallothionein.
Biochemistry 27:  8509-8515, 1988.

Hamer, D.H. Metallothionein.  Ann. Rev. Biochemistry 55: 913-951, 1986.

Bauman, J.W., Liu, J., Liu, Y.P., and Klaassen, C.D. Increase in
Metallothionein produced by chemicals that induce oxidative stress.
Toxicol.  Appl.  Pharmacol.  110: 347-354, 1991.

Cousins, R.J. Absorption, transport and hepatic metabolism of copper
and zinc:  Special reference to metallothionein and ceruloplasmin.
Physiol.  Rev. 65:  238-309, 1985.

Feldman, S.L., Failla, M.L., and Cousins, R.J. Degradation of liver
metallothioneins in vitro.  Biochem.  Biophys. Acta 544:  638-646,
1978.

Roesijadi, G. Metallothioneins in metal regulation and toxicity in
aquatic animals.  Aquatic Toxicology 22: 81-114, 1992.

Bremner, I. Nutritional and physiological significance of
metallothionein.  In:  J.H. R. Kaji (ed.), Metallothionein II, pp.
81-107.  Basel:  Birkhauser, 1987.

Chernaik, M.L. and Huang, P.C. Differential effect of
cysteine-to-serine substitutions in metallothionein on cadmium
resistance.  Proc.  Natl.  Acad.  Sci. USA 88: 3024-3028, 1991.

Waalkes, M.P. and Ward, J.M. Induction of hepatic metallothionein in
male B6C3F1 mice exposed to hepatic tumor promoters:  Effect on
phenobarbital, acetaminophen, sodium barbital, and Di(2-ethylhexyl)
phthalate.  Toxicol.  Appl. Pharmacol.  100:  217-226, 1989.

Kontozoglou, T.E., Banerjee, D., and Cherian, M.G. Immunohistochemical
localization of metallothionein in human testicular embryonal carcinoma
cells.  Virchows Archiv.  A Pathol.  Anat.  415:  545-549, 1989.

Muller, T., Schuckelt, R., and Jaenicke, L.
Cadmium/zinc-metallothionein induces DNA strand breaks in vitro.  Arch.
Toxicol.  65:  20-26, 1991.

Huang, P.C., Cody, C., Cismowski, M., Chernaik, M., Rhee, I.K., and
Lin, L.Y. Native and engineered metallothioneins. In:  C. Klaassen, and
K. T. Suzuki (eds.), Metallothionein in Medicine and Biology, pp.
87-101.  Florida:  CRC Press, 1991.

Zeng, J., Vallee, B.L., and Kagi, J.H.R. Zinc transfer from
transcription factor IIIA fingers to thionein clusters. Proc.  Natl.
Acad.  Sci. USA 88:  9984-9988, 1991.

Waalkes, M.P., Rehm, S., Riggs, C.W., Bare, R.M., Devor, D.E., Poirier,
L.A., Wenk, M.L., Henneman, J.R., and Balaschak, M.S. Cadmium
carcinogenesis in male Wistar[Crl:(W1)BR]rats:  Dose-response analysis
of tumor induction in the prostate and testes and at the injection
site.  Cancer Research 48:  4656-4663, 1988.

Kelley, S.L., Basu, A., Teicher, B.A., Hacker, M.P., Hamer, D.H., and
Lazo, J.S. Overexpression of metallothionein confers resistance to
anticancer drugs.  Science 241: 1813-1815, 1988.

Kasahara, K., Fujiwara, Y., Nishio, K., Ohmori, T., Sugimoto, Y.,
Komiya, K., Matsuda, T., and Saijo, N. Metallothionein content
correlates with the sensitivity of human small cell lung cancer cells
lines to cisplatin. Cancer Research 51:  3237-3242, 1991.