DIET, DNA METHYLATION AND OTHER EPIGENETIC EVENTS, AND CANCER PREVENTION
RELEASE DATE: September 27, 2002
RFA: CA-03-016
National Cancer Institute (NCI)
(http://www.nci.nih.gov/)
The Office of Dietary Supplements (ODS)
(http://www.nih.gov/icd/od)
LETTER OF INTENT RECEIPT DATE: February 18, 2003
APPLICATION RECEIPT DATE: March 18, 2003
THIS RFA CONTAINS THE FOLLOWING INFORMATION
o Purpose of this RFA
o Research Objectives
o Mechanism of Support
o Funds Available
o Eligible Institutions
o Individuals Eligible to Become Principal Investigators
o Where to Send Inquiries
o Letter of Intent
o Submitting an Application
o Peer Review Process
o Review Criteria
o Receipt and Review Schedule
o Award Criteria
o Required Federal Citations
PURPOSE
The NCI and the ODS invite applications for new R01 and R21 grants which are
focused on research leading to the elucidation of mechanism(s) by which
dietary factors influence epigenetic processes as well as increasing the
understanding of these processes in cancer prevention. The approach is to
encourage collaboration between nutrition and epigenetic /DNA methylation
experts to study bioactive food components with cancer preventative
properties, and to examine key epigenetic events in cancer processes (i.e.,
carcinogen metabolism, cell division, differentiation, apoptosis) so that
investigators can begin to establish linkages between epigenetics, methylation
pattern, and tumor incidence/behavior. It is anticipated that the information
gained will provide guidance for the development of dietary intervention
strategies for cancer prevention.
In addition to the present announcement, the NCI and the ODS announce a
related initiative about diet, DNA methylation and other epigenetic events,
and cancer prevention utilizing an additional funding mechanism: Competing
Supplements for existing NCI awardees
(https://grants.nih.gov/grants/guide/pa-files/PAR-02-175.html)
which can extend the goals of active grants to include studies related to the
impact of diet and nutrition on epigenetic events. Contact individuals listed
under "INQUIRIES" of this RFA for further information.
RESEARCH OBJECTIVES
Background
Cancer is a manifestation of both abnormal genetic and epigenetic events. The
importance of epigenetic events is that it represents a mechanism by which gene
function is selectively activated or inactivated. Actually DNA methylation is the
covalent addition of a methyl group to the 5 position of cytosine within CpG
dinucleotides and is a fundamental process that not only modulates gene expression,
but is also key to regulating chromosomal stability. A variety of regulatory
proteins including DNA methyltransferases, methyl-CpG binding proteins, histone-
modifying enzymes, chromatin remodeling factors, and their multimolecular complexes
are involved in the overall epigenetic process. Since epigenetic events are
susceptible to change they represent excellent targets to explain how environmental
factors, including diet, may modify cancer risk and tumor behavior. Abnormal DNA
methylation patterns are a hallmark of most cancers, including those of high
proportion in the United States i.e., colon, lung, prostate, and breast cancer.
Preclinical and clinical studies provide intriguing evidence that part of the
anticancer properties attributed to several bioactive food components,
encompassing both essential and non-essential nutrients, may relate to DNA
methylation patterns. There are four ways in which nutrients may be
interrelated with DNA methylation. The first is that nutrients may influence
the supply of methyl groups for the formation of S-adenosylmethionine (SAM).
The second mechanism is that nutrients may modify utilization of methyl groups
by processes including shifts in DNA methyltransferase activity. A third
plausible mechanism may relate to DNA demethylation activity. Finally, the
DNA methylation patterns may influence the response to a nutrient. These
interactions are described in greater detail below.
Global DNA Methylation Patterns
Global hypomethylation, accompanied by region-specific hypermethylation, is a
common characteristic among tumor cells. Global genomic hypomethylation has
been linked to the induction of chromosomal instability. Hypermethylation is
associated with the inactivation of virtually all pathways involved with the
cancer process, including DNA repair (e.g., hMLH1, BRCA1, MGMT), cell cycle
regulation (e.g., p16(INK4a), PTEN), inflammatory/stress response (e.g., COX-
2) and apoptosis (e.g., DAPK, APAF-1). Thus, evidence exists that variations
in the degree or site of DNA methylation can lead to abnormal DNA repair and
influence multiple cancer related genes and thereby influence the incidence
and behavior of tumors.
Stress including that resulting from dietary methionine/choline, folate, zinc,
and selenium inadequacy, as well as excessive alcohol intake can lead to
global DNA hypomethylation. Interestingly, the continuous feeding of a diet
deficient in choline and methionine is recognized to lead to global DNA
hypomethylation and cause hepatocellular carcinomas in rats in the absence of
any exogenous carcinogen. Paradoxically, either deficient or excess dietary
arsenic has also been shown to lead to global hypomethylation in rat liver.
Treatment with excess retinoic acid can also lead to hypomethylation as
recently shown in rat liver. Clinically, global DNA hypomethylation has been
observed in lymphocytic DNA from individuals consuming inadequate folate.
While limitations in the supply of methyl groups appear to be a common
mechanism, available data suggest that other factors determining DNA
methylation, including DNA methyltransferase (Dnmt), may be influenced by
bioactive food components.
Availability and Utilization of Methyl Groups
SAM and S-adenosylhomocysteine (SAH), as components of methyl metabolism, are
the substrate and product of essential cellular methyltransferase reactions.
SAM is derived from an ATP-dependent transfer of adenosine to methionine via
methionine adenosyltransferase and serves as the proximal methyl donor for
most methylation reactions. Cellular methyl acceptors include phospholipids,
proteins, histones, neurotransmitters, RNA and DNA. The formation of SAM
requires a continuous supply of folate, methionine, choline, vitamin B12,
vitamin B6, vitamin B2 and energy from the extracellular milieu.
Polymorphisms in the methyl metabolism genes methionine synthase,
methylenetetrahydrofolate reductase (MTHFR), and cystathione beta-synthetase
affect concentrations of SAM and likely modify the susceptibility to cancer.
Investigators are currently exploring the interaction between these
polymorphisms, diet and DNA methylation. For example, subjects homozygous for
the C677T polymorphism in the MTHFR gene exhibited a significantly lower level
of methylated DNA but only under conditions of low folate status.
The Dnmts are a family of enzymes that catalyze the transfer of methyl groups
from SAM to cytosine residues in DNA. This produces 5-methylcytosine and SAH.
Higher DNA methyltransferase activity has been observed in tumor cells
compared to normal cells. Chronic dietary methyl deficiency also increases
DNA methyltransferase activity, which may be an attempt to compensate for
diminished SAM supply. Nutrient supply, therefore, appears to play a key role
in regulating DNA methyltransferase activity.
Selenium is a dietary trace element that is recognized as having potential
anticancer properties. Interest in selenium as a prostate cancer preventative
nutrient is showcased by the recent initiation of the SELECT trial by NCI
(http://newscenter.cancer.gov/pressreleases/SELECT.html). While selenium is
known to modify several aspects of the cancer process, the one that is most
critical for bringing about a phenotypic change remains to be established.
Interestingly, increased selenium concentrations have been found to inhibit
Dnmt1 activity in vitro and decrease Dnmt1 protein expression in vitro.
Consistent with these data, selenium deficiency leads to increased Dnmt1
protein expression in vitro. The effects of selenium on Dnmt activity suggest
that dietary factors influencing methyl utilization may also modify DNA
methylation patterns.
Although the enzymes for direct methylation of DNA are well characterized, the
demethylation process has been assumed to be a passive process. A mammalian
gene that codes for a protein that can catalyze demethylation recently has
been described, however other attempts have yielded variable results.
Demethylation activity has been hypothesized to be higher in non-neoplastic
compared to neoplastic cells. Loss of demethylation activity may account for
some of the differences in methylation patterns between neoplastic and normal
tissue. The role, if any, of bioactive food components in regulating the
demethylation process remains to be established.
Gene-Specific DNA Methylation Patterns
Fluctuations in the degree of CpG island methylation are key to regulating
functional promoters by modifying the binding of transcription factors and
methyl-DNA binding proteins. Aberrant methylation of CpG islands on the
promoter region may contribute to the progressive inactivation of growth-
inhibitory genes resulting in the clonal selection of cells with growth
advantage during cancer development.
While methyl-deficiency leads to global DNA hypomethylation, it also
simultaneously leads to gene-specific hypomethylation and hypermethylation.
Preclinical studies reveal that consumption of a methyl deficient diet leads
to hypomethylation of specific CpG sites within several genes including c-myc,
c-fos and c-H-ras. This hypomethylation was accompanied by elevated levels of
mRNA for these same genes. Folate depletion was shown to produce gene-
specific, rather than global DNA hypomethylation in human nasopharyngeal
carcinoma KB cells. Interestingly, these investigators also reported that
folate depletion led to hypermethylation of a 5" sequence of the H-cadherin
gene, which accompanied diminished mRNA expression. These data suggest that
global DNA hypomethylation may incompletely predict the response to an
individual dietary component. More probing studies are needed to characterize
gene-specific changes brought about by bioactive food components.
Epigenetic events occurring in utero can lead to persistent changes in gene
expression and can be modified by the diet. For example, the diet provided to
female mice of two strains during pregnancy modified the offsprings" hair coat
color (increased agouti vs yellow) and DNA methylation patterns. Expression
of the yellow hair color in these mice is recognized to be controlled by
hypomethylation of the agouti long terminal repeat (LTR), which was
hypermethylated by dietary supplementation of the maternal diet with zinc,
methionine, betaine, choline, folate and vitamin B12. Expression of this
yellow coat color is linked with increased risk of obesity, diabetes, and
cancer. While long-term health implications remain to be determined, these
studies demonstrate that feeding a methyl-supplemented diet increased levels
of DNA methylation in the agouti LTR and increased the proportion of agouti to
yellow mice. It should be noted that this effect occurred with control diets
that are considered adequate for meeting nutritional needs. These data point
to the likelihood that in utero nutrient exposures can lead to genomic
imprinting in the offspring and potentially modify cancer risk.
Invariably, increased fruit and vegetable consumption is associated with
reduction in cancer risk. Evidence from a variety of sources suggests that
flavonoids, including genistein may have merit as an effective deterrent of
cancer. Dietary genistein has been shown to lead to changes in DNA
methylation patterns in prostate tissue of C57BL/6J mice. After sequencing,
it was determined that genistein consumption led to hypermethylation at 4
specific CpG islands, including a dexamethasone-induced product gene (D44443).
Neonatal exposure to the phytoestrogens coumestrol and equol has been found to
lead to specific gene hypermethylation in the c-H-ras proto-oncogene in
pancreatic DNA. Data indicating that consumption of high fiber diets is
accompanied by a reduction in estrogen receptor methylation in colon tissue
from healthy subjects has also been noted. Hypermethylation of the promoter
CpG islands is recognized to contribute to the loss of function of several
tumor related genes, including estrogen receptor (ER). Overall, several
studies illustrate that bioactive food components other than essential
nutrients can influence DNA methylation processes.
Cell Responsiveness to DNA Methylation Defects
The ability of bioactive food components to reduce proliferation in normal
cells has been reported to be less than in neoplastic cells. For example,
selenium preferentially induces growth inhibition and apoptosis in prostate
cancer cells, but not in normal prostate cells. Recently, the flavonoid
apigenin has also shown selective growth inhibition in prostate cancer cells
without affecting normal cells. It is unclear if these differences in
sensitivity relate to epigenetic and DNA methylation processes.
The gene encoding the pi class glutathione S-transferase (GSTP1) is known to
be silenced by CpG island methylation in prostate cancer cells. It has been
suggested that compensation for loss of GSTP1 activity is possible with
inducers of general glutathione S-transferase activity and one such inducer is
the isothiocyanate compound sulforophane found in cruciferous vegetables. That
dietary factors might circumvent changes induced by aberrant DNA methylation
patterns in cancer requires investigation.
The short-chain fatty acid butyrate induces cell cycle arrest, differentiation
and apoptosis in colon cancer cells, but often induces opposite effects in
normal colonic epithelial cells. A stable transgenic cell line containing a
lacZ reporter gene coupled with a hormone-inducible promoter was examined for
reversal of methylation-mediated gene silencing. In the methylated form, the
promoter is unable to induce lacZ expression and is insensitive to
reactivation by the histone deacetylase inhibitors trichostatin A and
suberoylanilide hydroxamic acid. In contrast, the short-chain fatty acids
(including proprionic, butyric and valeric acid) were capable of overcoming
methylation-mediated repression and restored lacZ expression to levels
approaching that from an unmethylated promoter.
Retinoic acid provides an example that some nutrients may also restore
methylation patterns and gene expression. RA demethylated the retinoic acid
receptor beta2 promoter in acute promyeocytic leukemia cells. This led to
cellular differentiation and accumulation of retinoic acid receptor beta2
mRNA in these cells. It is unknown whether other bioactive food components
also lead to DNA demethylation.
Sensitizing tumor cells to the bioactive food component can also be
accomplished by using demethylating agents, such as 5-aza-2"-deoxycytidine (5-
aza-dC). This compound has been used as a therapeutic agent for certain
cancers, although treatment-associated side effects preclude its general
usefulness. Treatment of several cell lines with MLL (mixed-lineage leukemia
or myeloid/lymphoid leukemia) translocations with 5-aza-dC increases the
ability of all-trans retinoic acid or 1,25-dihydroxyvitamin D3 to cause
differentiation. This study suggests that methylation patterns can influence
the response to bioactive food components. These are intriguing results, but
we need more.
Other Epigenetic Events
Emerging evidence indicates that various chromatin states such as histone
modifications (acetylation and methylation) and nucleosome positioning
(modulated by ATP-dependent chromatin remodeling machines) determine DNA
methylation patterning. Additionally, various regulatory factors interacting
with the DNA methyltransferases may direct them to specific DNA sequences,
regulate their enzymatic activity, and allow their use as transcriptional
repressors. Continued studies of the connections between DNA methylation and
chromatin structure and the DNA methyltransferase-associated proteins, will
likely reveal that many epigenetic modifications of the genome are directly
connected. DNA hypermethylation is not likely to be an isolated layer of
epigenetic control, but is likely to be linked to other pieces of the puzzle
such as methyl-binding proteins, DNA methyltransferases and histone
deacetylase. Our understanding of the degree of specificity of these
epigenetic layers in the control of gene function remains incomplete.
Histone modifications have recently generated excitement in epigenetic
research, culminating in the `histone code" hypothesis. The idea is that the
modified histone tails provide binding sites for chromatin-associated
proteins, which in turn induce alterations in chromatin structure and other
downstream events. In particular, for silent chromatin domains, histone H3
lysine 9 methylation by the Su(var)3-9 family of histone methyltransferases
has been shown to play a key part in the formation of transcriptionally
repressed chromatin by providing a high-affinity binding site for the
chromodomain of the heterochromatic HP1 proteins. Recent evidence suggests
that histone methylation has considerable epigenetic regulatory impact and is
likely influenced by dietary methyl group sufficiency. Moreover, the recent
finding that some histone methyltransferases function as tumor suppressors
provides another rationale for continued exploration of the diet and
epigenetic relationship in cancer prevention research.
Acetylation of core histone tails is a conserved mechanism modulating gene
expression. It is able to relax higher order chromatin, to promote factor
binding to DNA and to recruit unique biological complexes that mediate
downstream functions. A switch from inactive to active chromatin is often
accompanied by histone hyperacetylation of critical sites in gene regulatory
regions. The antagonistic activities of histone acetyltransferases and histone
deacetylases control the nuclear steady-state balance of this covalent
modification. In addition to any effect on methylation patterns, the short-
chain fatty acid butyrate has been shown to induce histone hyperacetylation in
vitro and inhibit histone deacetylase in vitro. Recent evidence demonstrates
that butyrate promotes hyperacetylation of the upstream region of the RET gene
(a proto-oncogene which encodes a tyrosine kinase receptor predominantly
expressed in the developing embryo) and increases transcription of the gene.
More probing studies are needed to characterize additional gene-specific
acetylation changes brought about by bioactive food components.
Calorie restriction is one of the best-documented and most potent experimental
manipulations for decreasing tumor development in rodents and increasing
longevity in diverse organisms. The mechanisms underlying the antitumorigenic
effects of caloric restriction have not yet been elucidated. Nevertheless,
there is great interest in translating the anticarcinogenic effects of caloric
restriction into prevention strategies. Recent observations suggest that a
protein important for heterochromatin formation, Sir2, is central for caloric-
restriction induced longevity in animals. Any role for Sir2 in cancer
prevention requires elucidation.
Objectives and Scope
This RFA seeks to promote innovative, preclinical and clinical research to
determine how diet and dietary factors impact DNA methylation and other
epigenetic processes involved with cancer prevention. Although much evidence
exists that dietary components are linked to cancer prevention, the specific
nutrients and sites of action remain elusive. Diet, in fact, has been
implicated in many of the pathways of cancer, including apoptosis, cell cycle
control, differentiation, inflammation, angiogenesis, DNA repair and
carcinogen metabolism. These are also processes that are likely regulated by
DNA methylation, and possibly other epigenetic events, which impact gene
function. The objective of this RFA is to begin to address the following
issues: how bioactive food components regulate DNA methylation or other
epigenetic events for cancer prevention, if bioactive food components can
alter DNA methylation or other aberrant epigenetic events and restore gene
function, and if these components can circumvent or compensate for genes and
pathways that are altered by epigenetic events.
An important aim of this RFA is to encourage collaboration between nutrition
and epigenetic/ DNA methylation experts to study bioactive food components
with cancer preventative properties and to examine key epigenetic events in
cancer processes (i.e., carcinogen metabolism, cell division, differentiation,
apoptosis) in order to begin to establish linkages between epigenetics,
methylation pattern, and tumor incidence/behavior. Thus, each application
should demonstrate experience in nutrition and cancer prevention as well as
DNA methylation or epigenetics.
Studies should also link phenotypic changes to epigenetic alterations induced
by specific essential and non-essential nutrients. The resulting information
will be critical for optimizing effective dietary intervention strategies for
cancer prevention. Investigators may choose from the full range of clinical
and preclinical approaches. The focus should be on how individual dietary
components influence DNA methylation and other epigenetic events and how this
correlates with phenotypic change. Very little information currently exists
about gene-specific changes in DNA methylation as influenced by bioactive food
components, furthermore, very little information exists to adequately evaluate
the specificity of individual nutrients, the impact of intakes/exposures, and
any acclimation with time and/or tissue specificity. This RFA encourages the
submission of novel approaches to unravel relationships between DNA
methylation and other epigenetic events, diet, and cancer prevention.
A variety of technologies to assess DNA methylation sequences may be utilized.
These can be broadly classified into techniques that measure the overall
content or distribution patterns of 5-methylcytosine (i.e., methylated CpG
island amplification, methylation-sensitive restriction fingerprinting,
differential methylation hybridization, and Restriction Landmark Genomic
Scanning) and those that examine known gene sequences (i.e., methylation-
sensitive single nucleotide primer extension, and combined bisulfite
restriction analysis). The use of genetically engineered animal models
including transgenic or gene knockouts are appropriate. The efficient
utilization of molecular resources such as gene databases and bioinformatics
may also be used to expedite identification of gene-specific methylation
targets.
The following are viewed as relevant examples for the development of the R01
and R21 applications:
o Gene-specific changes in DNA methylation caused by excesses and limitations
in bioactive food components.
o Relationship between dietary induced global DNA hypomethylation and gene-
specific hypermethylation.
o Relationship between diet induced changes in DNA methylation and histone
acetylation/methylation and control of gene function
o Temporality in DNA methylation patterns as influenced by bioactive food
components.
o Bioactive food component regulation of DNA methylation/epigenetic processes,
i.e., methylenetetrahydrofolate reductase, methionine synthase, DNA
methyltransferases, demethylases (or various demethylation processes),
methylcytosine-binding proteins, histone methyltransferases, histone
aceyltransferases and deacetylases.
o Linkages between DNA methylation or other epigenetic pattern and the
anticancer properties of bioactive food components.
MECHANISM OF SUPPORT
This RFA will use the National Institutes of Health (NIH) research project
grant (R01) and exploratory/developmental grants (R21) as award mechanisms.
As an applicant you will be solely responsible for planning, directing, and
executing the proposed project. Future unsolicited, competing-continuation
applications based on this project will compete with all investigator-
initiated applications and will be reviewed according to the customary peer
review procedures. The anticipated award date is December 2003.
This RFA uses just-in-time concepts. It also uses the modular as well as the
non-modular budgeting formats (see
https://grants.nih.gov/grants/funding/modular/modular.htm). Specifically, if
you are submitting an application with direct costs in each year of $250,000
or less, use the modular format. Otherwise follow the instructions for non-
modular research grant applications.
FUNDS AVAILABLE
NCI intends to commit approximately $2.5 million in FY 2004 to fund 7 to 10
grants in response to this RFA. The ODS intends to commit up to $800,000 in
FY 2004 to fund 3 to 4 grants in response to this RFA. An R01 applicant may
request a project period of up to 4 years. An R21 applicant may request a
project period of up to 2 years and a budget for direct costs of up to
$100,000 per year. Because the nature and scope of the proposed research will
vary from application to application, it is anticipated that the size and
duration of each award will also vary. Although the financial plans of NCI and
the ODS provide support for this program, awards pursuant to this RFA are
contingent upon the availability of funds and the receipt of a sufficient
number of meritorious applications. At this time, it is not known if this RFA
will be reissued.
ELIGIBLE INSTITUTIONS
You may submit (an) application(s) if your institution has any of the
following characteristics:
o For-profit or non-profit organizations
o Public or private institutions, such as universities, colleges, hospitals,
and laboratories
o Units of State and local governments
o Eligible agencies of the Federal government
o Domestic or foreign
o Faith-based or community based organizations
INDIVIDUALS ELIGIBLE TO BECOME PRINCIPAL INVESTIGATORS
Any individual with the skills, knowledge, and resources necessary to carry
out the proposed research is invited to work with their institution to develop
an application for support. Individuals from underrepresented racial and
ethnic groups as well as individuals with disabilities are always encouraged
to apply for NIH programs.
WHERE TO SEND INQUIRIES
We encourage inquiries concerning this RFA and welcome the opportunity to
answer questions from potential applicants. Inquiries may fall into three
areas: scientific/research, peer review, and financial or grants management
issues:
o Direct your questions about scientific/research issues related to diet and
cancer prevention to:
Dr. Sharon A. Ross
Division of Cancer Prevention
National Cancer Institute
6130 Executive Blvd., Room 3157
Bethesda, MD 20892
Telephone: (301) 594-7547
FAX: (301) 480-3925
Email: sr75k@nih.gov
o Direct your questions about scientific/research issues related to dietary
supplements to:
Rebecca B. Costello, Ph.D.
Office of Dietary Supplements
Office of Disease Prevention, Office of the Director
National Institutes of Health
6100 Executive Blvd., Room 3B01, MSC 7517
Bethesda, Maryland 20892-7517
Phone: 301-435-2920
Fax: 301-480-1845
Email: CostellB@od.nih.gov
o Direct your questions about peer review issues to:
Referral Officer
National Cancer Institute
Division of Extramural Activities
6116 Executive Boulevard, Room 8041, MSC 8329
Bethesda, MD 20892-8329
Telephone: (301) 496-3428
FAX: (301) 402-0275
Email: ncidearefof@mail.nih.gov
o Direct your questions about financial or grants management matters to:
Jane Paull
Grants Administration Branch
National Cancer Institute
6120 Executive Blvd., EPS-243
Bethesda, MD 20892
Telephone: (301) 496-2182
FAX: (301) 496-8601
Email: paullj.gab.nci.nih.gov
LETTER OF INTENT
Prospective applicants are asked to submit a letter of intent that includes
the following information:
o Descriptive title of the proposed research
o Name, address, and telephone number of the Principal Investigator
o Names of other key personnel
o Participating institutions
o Number and title of this RFA
Although a letter of intent is not required, is not binding, and does not
enter into the review of a subsequent application, the information that it
contains allows NCI staff to estimate the potential review workload and plan
the review.
The letter of intent is to be sent by the date listed at the beginning of this
document. The letter of intent should be sent to:
Dr. Sharon A. Ross
Division of Cancer Prevention
National Cancer Institute
6130 Executive Blvd., Room 3157
Bethesda, MD 20892
Telephone: (301) 594-7547
FAX: (301) 480-3925
Email: sr75k@nih.gov
SUBMITTING AN APPLICATION
Applications must be prepared using the PHS 398 research grant application
instructions and forms (rev. 5/2001). The PHS 398 is available at
https://grants.nih.gov/grants/funding/phs398/phs398.html in an interactive
format. For further assistance contact GrantsInfo, Telephone (301) 710-0267,
Email: GrantsInfo@nih.gov.
SPECIFIC INSTRUCTIONS FOR MODULAR GRANT APPLICATIONS: Applications requesting
up to $250,000 per year in direct costs must be submitted in a modular grant
format. The modular grant format simplifies the preparation of the budget in
these applications by limiting the level of budgetary detail. Applicants
request direct costs in $25,000 modules. Section C of the research grant
application instructions for the PHS 398 (rev. 5/2001) at
https://grants.nih.gov/grants/funding/phs398/phs398.html includes step-by-step
guidance for preparing modular grants. Additional information on modular
grants is available at
https://grants.nih.gov/grants/funding/modular/modular.htm.
USING THE RFA LABEL: The RFA label available in the PHS 398 (rev. 5/2001)
application form must be affixed to the bottom of the face page of the
application. Type the RFA number on the label. 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 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 RFA label is also available at:
https://grants.nih.gov/grants/funding/phs398/label-bk.pdf.
All application instructions outlined in the PHS 398 application kit are to be
followed, with the following requirements for R21 applications:
1. R21 applications will use the "MODULAR GRANT" and "JUST-IN-TIME" concepts,
with direct costs requested in $25,000 modules, up to the total direct costs
limit of $100,000 per year.
2. Although preliminary data are not required for an R21 application, they may
be included.
3. Sections a-d of the Research Plan of the R21 application may not exceed 25
pages, including tables and figures.
4. R21 appendix materials should be limited, as is consistent with the
exploratory nature of the R21 mechanism, and should not be used to circumvent
the page limit for the research plan. Copies of appendix material will only
be provided to the primary reviewers of the application and will not be
reproduced for wider distribution. The following materials may be included in
the appendix:
o Up to five publications, including manuscripts (submitted or accepted for
publication), abstracts, patents, or other printed materials directly relevant
to the project. These may be stapled as sets. Surveys, questionnaires, data
collection instruments, and clinical protocols. These may be stapled as sets.
Original glossy photographs or color images of gels, micrographs, etc.,
provided that a photocopy (may be reduced in size) is also included within the
25 page limit of items a-d of the research plan.
SENDING AN APPLICATION TO THE NIH: Submit a signed, typewritten original of
the application, including the Checklist, and three signed, photocopies, in
one package 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)
At the time of submission, two additional copies of the application must be
sent to:
Referral Officer
Division of Extramural Activities
National Cancer Institute
6116 Executive Blvd., Room 8041, MSC-8329
Rockville, MD 20852 (express courier)
Bethesda MD 20892-8329
APPLICATIONS HAND-DELIVERED BY INDIVIDUALS TO THE NATIONAL CANCER INSTITUTE
WILL NO LONGER BE ACCEPTED. This policy does not apply to courier deliveries
(i.e. FEDEX, UPS, DHL, etc.)
(https://grants.nih.gov/grants/guide/notice-files/NOT-CA-02-002.html)
This policy is similar to and consistent with the policy for applications
addressed to Centers for Scientific Review as published in the NIH Guide Notice
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-02-012.html.
APPLICATION PROCESSING: Applications must be received by the application
receipt date listed in the heading of this RFA. If an application is received
after that date, it will be returned to the applicant without review.
The Center for Scientific Review (CSR) will not accept any application in
response to this RFA that is essentially the same as one currently pending
initial review, unless the applicant withdraws the pending application. The
CSR 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.
PEER REVIEW PROCESS
Upon receipt, applications will be reviewed for completeness by the CSR and
for responsiveness by the NCI and ODS program staff.
Incomplete applications will be returned to the applicant without further
consideration. And, if the application is not responsive to the RFA, CSR
staff may contact the applicant to determine whether to return the application
to the applicant or submit it for review in competition with unsolicited
applications at the next appropriate NIH review cycle.
Applications that are complete and responsive to the RFA will be evaluated for
scientific and technical merit by an appropriate peer review group convened by
the Division of Extramural Activities (DEA) at NCI in accordance with the
review criteria stated below. As part of the initial merit review, all
applications will:
o Receive a written critique
o Undergo a process in which only those applications deemed to have the
highest scientific merit, generally the top half of the applications under
review, will be discussed and assigned a priority score
o Those that receive a priority score will undergo a second level review by
the National Cancer Advisory 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. In
the written comments, reviewers will be asked to discuss the following aspects
of your application in order to judge the likelihood that the proposed
research will have a substantial impact on the pursuit of these goals:
o Significance
o Approach
o Innovation
o Investigator
o Environment
The scientific review group will address and consider each of these criteria
in assigning your application"s overall score, weighting them as appropriate
for each application. Your application does not need to be strong in all
categories to be judged likely to have major scientific impact and thus
deserve a high priority score. For example, you 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 question about the
role of (a) bioactive food components in DNA methylation or other epigenetic
event involved with cell vulnerability to cancer or cellular responsiveness to
cancer prevention? Do studies focus on how dietary components influence DNA
methylation or other epigenetic event and how this correlates with phenotypic
change. Will these research projects advance the field of nutrition and
cancer prevention from observational to more probing studies?
(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
methods? 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 experience in nutrition and cancer prevention as
well as DNA methylation or epigenetics demonstrated in the application? 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 (i.e., between nutrition and epigenetic /DNA
methylation experts)? Is there evidence of institutional support?
ADDITIONAL REVIEW CRITERIA: In addition to the above criteria, your
application will also be reviewed with respect to the following:
o PROTECTIONS: The adequacy of the proposed protection for humans, animals,
or the environment, to the extent they may be adversely affected by the
project proposed in the application.
o INCLUSION: The adequacy of plans to include subjects from both genders, all
racial and ethnic groups (and subgroups), and children as appropriate for the
scientific goals of the research. Plans for the recruitment and retention of
subjects will also be evaluated. (See Inclusion Criteria included in the
section on Federal Citations, below)
o DATA SHARING: The adequacy of the proposed plan to share data.
o BUDGET: The reasonableness of the proposed budget and the requested period
of support in relation to the proposed research.
RECEIPT AND REVIEW SCHEDULE
Letter of Intent Receipt Date: February 18, 2003
Application Receipt Date: March 18, 2003
Peer Review Date: June-July, 2003
Council Review: September 2003
Earliest Anticipated Start Date: December 2003
AWARD CRITERIA
Award criteria that will be used to make award decisions include:
o Scientific merit (as determined by peer review)
o Availability of funds
o Programmatic priorities.
REQUIRED FEDERAL CITATIONS
MONITORING PLAN AND DATA SAFETY AND MONITORING BOARD: Research components
involving Phase I and II clinical trials must include provisions for
assessment of patient eligibility and status, rigorous data management,
quality assurance, and auditing procedures. In addition, it is NIH policy
that all clinical trials require data and safety monitoring, with the method
and degree of monitoring being commensurate with the risks (NIH Policy for
Data Safety and Monitoring, NIH Guide for Grants and Contracts, June 12, 1998:
https://grants.nih.gov/grants/guide/notice-files/not98-084.html).
Clinical trials supported or performed by NCI require special considerations.
The method and degree of monitoring should be commensurate with the degree of
risk involved in participation and the size and complexity of the clinical
trial. Monitoring exists on a continuum from monitoring by the principal
investigator/project manager or NCI program staff or a Data and Safety
Monitoring Board (DSMB). These monitoring activities are distinct from the
requirement for study review and approval by an Institutional review Board
(IRB). For details about the Policy for the NCI for Data and Safety
Monitoring of Clinical trials see:
http://deainfo.nci.nih.gov/grantspolicies/datasafety.htm. For Phase I and II
clinical trials, investigators must submit a general description of the data
and safety monitoring plan as part of the research application. See NIH Guide
Notice on "Further Guidance on a Data and Safety Monitoring for Phase I and II
Trials" for additional information:
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-00-038.html.
Information concerning essential elements of data safety monitoring plans for
clinical trials funded by the NCI is available:
http://www.cancer.gov/clinical_trials/
INCLUSION OF WOMEN AND MINORITIES IN CLINICAL RESEARCH: 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 clinical research projects unless a
clear and compelling justification is provided indicating 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 clinical research should read the AMENDMENT "NIH
Guidelines for Inclusion of Women and Minorities as Subjects in Clinical
Research - Amended, October, 2001," published in the NIH Guide for Grants and
Contracts on October 9, 2001
(https://grants.nih.gov/grants/guide/notice-files/NOT-OD-02-001.html),
a complete copy of the updated Guidelines are available at
https://grants.nih.gov/grants/funding/women_min/guidelines_amended_10_2001.htm.
The amended policy incorporates: the use of an NIH definition of
clinical research, updated racial and ethnic categories in compliance with the
new OMB standards, clarification of language governing NIH-defined Phase III
clinical trials consistent with the new PHS Form 398, and updated roles and
responsibilities of NIH staff and the extramural community. The policy
continues to require for all NIH-defined Phase III clinical trials that: a)
all applications or proposals and/or protocols must provide a description of
plans to conduct analyses, as appropriate, to address differences by
sex/gender and/or racial/ethnic groups, including subgroups if applicable, and
b) investigators must report annual accrual and progress in conducting
analyses, as appropriate, by sex/gender and/or racial/ethnic group
differences.
INCLUSION OF CHILDREN AS PARTICIPANTS IN RESEARCH INVOLVING HUMAN SUBJECTS:
The NIH maintains a policy 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 is available at
https://grants.nih.gov/grants/funding/children/children.htm.
REQUIRED EDUCATION ON THE PROTECTION OF HUMAN SUBJECT PARTICIPANTS: NIH policy
requires education on the protection of human subject participants for all
investigators submitting NIH proposals for research involving human subjects.
You will find this policy announcement in the NIH Guide for Grants and
Contracts Announcement, dated June 5, 2000, at
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-00-039.html. A
continuing education program in the protection of human participants in
research in now available online at: http://cme.nci.nih.gov/
HUMAN EMBRYONIC STEM CELLS (hESC): Criteria for federal funding of research on
hESCs can be found at https://grants.nih.gov/grants/stem_cells.htm and at
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-02-005.html. Guidance
for investigators and institutional review boards regarding research involving
human embryonic stem cells, germ cells, and stem cell-derived test articles
can be found at
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-02-044.html.
Only research using hESC lines that are registered in the NIH Human
Embryonic Stem Cell Registry will be eligible for Federal funding (see
http://escr.nih.gov). It is the responsibility of the applicant to provide
the official NIH identifier(s)for the hESC line(s) to be used in the proposed
research. Applications that do not provide this information will be returned
without review.
PUBLIC ACCESS TO RESEARCH DATA THROUGH THE FREEDOM OF INFORMATION ACT: The
Office of Management and Budget (OMB) Circular A-110 has been revised to
provide public access to research data through the Freedom of Information Act
(FOIA) under some circumstances. Data that are (1) first produced in a
project that is supported in whole or in part with Federal funds and (2) cited
publicly and officially by a Federal agency in support of an action that has
the force and effect of law (i.e., a regulation) may be accessed through FOIA.
It is important for applicants to understand the basic scope of this
amendment. NIH has provided guidance at
https://grants.nih.gov/grants/policy/a110/a110_guidance_dec1999.htm.
Applicants may wish to place data collected under this RFA in a public
archive, which can provide protections for the data and manage the
distribution for an indefinite period of time. If so, the application should
include a description of the archiving plan in the study design and include
information about this in the budget justification section of the application.
In addition, applicants should think about how to structure informed consent
statements and other human subjects procedures given the potential for wider
use of data collected under this award.
URLs IN NIH GRANT APPLICATIONS OR APPENDICES: All applications and proposals
for NIH funding must be self-contained within specified page limitations.
Unless otherwise specified in an NIH solicitation, Internet addresses (URLs)
should not be used to provide information necessary to the review because
reviewers are under no obligation to view the Internet sites. Furthermore,
we caution reviewers that their anonymity may be compromised when they
directly access an Internet site.
HEALTHY PEOPLE 2010: The Public Health Service (PHS) is committed to achieving
the health promotion and disease prevention objectives of "Healthy People
2010," a PHS-led national activity for setting priority areas. This RFA is
related to one or more of the priority areas. Potential applicants may obtain
a copy of "Healthy People 2010" at http://www.health.gov/healthypeople/
AUTHORITY AND REGULATIONS: This program is described in the Catalog of Federal
Domestic Assistance No. 93.3393 (Cancer Cause and Prevention Research) and is
not subject to the intergovernmental review requirements of Executive Order
12372 or Health Systems Agency review. Awards are made under authorization of
Sections 301 and 405 of the Public Health Service Act as amended (42 USC 241
and 284) and administered under NIH grants policies described at
https://grants.nih.gov/grants/policy/policy.htm and under Federal Regulations
42 CFR 52 and 45 CFR Parts 74 and 92.
The PHS strongly encourages all grant recipients to provide a smoke-free
workplace and discourage the 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.
The Office of Dietary Supplements (ODS) was mandated by Congress in 1994 and
established within the Office of the Director, National Institutes of Health
(NIH). The Dietary Supplement Health and Education Act (DSHEA) [Public Law
103-417, Section 3.a] amended the Federal Food, Drug, and Cosmetic Act "to
establish standards with respect to dietary supplements." This law authorized
the establishment of the ODS.