Expired PA-14-198: Unconventional Roles of Ethanol Metabolizing Enzymes, Metabolites, and Cofactors in Health and Disease (R01)

Part 1. Overview Information

Participating Organization(s)	National Institutes of Health (NIH)
Components of Participating Organizations	National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Funding Opportunity Title	Unconventional Roles of Ethanol Metabolizing Enzymes, Metabolites, and Cofactors in Health and Disease (R01)
Activity Code	R01 Research Project Grant
Announcement Type	Reissuance of PA-12-235
Related Notices	December 08, 2014 - This PA has been reissued as PA-15-058. June 4, 2014 - Notice NOT-14-074 supersedes instructions in Section III.3 regarding applications that are essentially the same.
Funding Opportunity Announcement (FOA) Number	PA-14-198
Companion Funding Opportunity	PA-12-234, R21 Exploratory/Developmental Grant PA-12-235, R01 Research Project Grant
Number of Applications	See Section III. 3. Additional Information on Eligibility.
Catalog of Federal Domestic Assistance (CFDA) Number(s)	93.273
Funding Opportunity Purpose	The purpose of this FOA is to provide support for integrated, innovative research on the novel and unconventional contributions of ethanol metabolizing pathways, their metabolites, cofactors, and interactions with synergizing biological pathways in the development of alcohol- induced diseases and end organ injuries. It is anticipated that this FOA will generate data that may lead to breakthroughs in our understanding of identifying key cellular and molecular components in the initiation, progression and maintenance of the diverse medical disorders caused by excessive, long term alcohol consumption. In the future this knowledge may be critical in the diagnosis, treatment and management of vulnerable patient population debilitated by the vast array of alcohol-induced pathologies and enable clinicians to improve disease outcomes and, consequently, public health.

Key Dates

Posted Date	May 1, 2014
Open Date (Earliest Submission Date)	May 2, 2014
Letter of Intent Due Date(s)	Not Applicable
Application Due Date(s)	Standard dates apply, by 5:00 PM local time of applicant organization. Applicants are encouraged to apply early to allow adequate time to make any corrections to errors found in the application during the submission process by the due date.
AIDS Application Due Date(s)	Standard AIDS dates apply, by 5:00 PM local time of applicant organization. Applicants are encouraged to apply early to allow adequate time to make any corrections to errors found in the application during the submission process by the due date.
Scientific Merit Review	Standard dates apply
Advisory Council Review	Standard dates apply
Earliest Start Date	Standard dates apply
Expiration Date	New Date: December 09, 2014. See issuance of PA-15-058. ~~January 8, 2015~~
Due Dates for E.O. 12372	Not Applicable

Required Application Instructions

It is critical that applicants follow the instructions in the SF424 (R&R) Application Guide, except where instructed to do otherwise (in this FOA or in a Notice from the NIH Guide for Grants and Contracts). Conformance to all requirements (both in the Application Guide and the FOA) is required and strictly enforced. Applicants must read and follow all application instructions in the Application Guide as well as any program-specific instructions noted in Section IV. When the program-specific instructions deviate from those in the Application Guide, follow the program-specific instructions. Applications that do not comply with these instructions may be delayed or not accepted for review.

Table of Contents

Part 1. Overview Information
Part 2. Full Text of the Announcement
Section I. Funding Opportunity Description
Section II. Award Information
Section III. Eligibility Information
Section IV. Application and Submission Information
Section V. Application Review Information
Section VI. Award Administration Information
Section VII. Agency Contacts
Section VIII. Other Information

Part 2. Full Text of Announcement

Section I. Funding Opportunity Description

Background

Alcohol is the most frequently abused drug for millennia and an etiological inducer of numerous human diseases. For many of these diseases, ethanol metabolism plays a key role. From a quantitative standpoint, ethanol is mainly metabolized in the liver, which is also the most severely affected organ by excessive alcohol consumption, however other organs metabolize ethanol as well to a much less extent. Organ specific diseases resulting from ethanol metabolism include alcoholic cardiomyopathy, leaky gut, neurodegeneration, kidney failure and alcoholic liver disease among others.

Alcohol also impairs liver’s capacity for activation, utilization of certain nutrients and vitamins and detoxification of chemicals. Unlike other drugs or abuse, alcohol is a source of energy that sometimes accounts for more than half of alcoholic’s caloric intake. As a result, it displaces normal nutrients, leading to malnutrition and vitamin deficiencies including folate, thiamin, vitamin A and others. Secondary malnutrition can result from deficient absorption and digestion of nutrients due to ineffective digestive enzymes and other gastrointestinal complications. Deranged liver metabolism and various ingestive behavior mechanisms including liver-brain-gut interactions further aggravate ethanol associated malnutrition.

The primary enzymes of alcohol metabolism are alcohol dehydrogenase (ADH), cytochrome p450 (CYP2E1) and, to a lesser extent, catalase, enzymes that primarily reside in the cytoplasm, the endoplasmic reticulum (ER) and the peroxisome, respectively. ADH and CYP2E1 constitute the two major pathways of ethanol oxidation and both produce highly reactive acetaldehyde. Acetaldehyde in turn is further converted to acetate by aldehyde dehydrogenases (ALDH). In addition to generating acetaldehyde, both reactions reduce NAD+ to NADH. Furthermore, ethanol metabolism by CYP2E1 also generates several reactive oxygen species (ROS) species. Acetaldehyde is highly toxic by forming protein and DNA adducts, direct inhibition of key enzymes leading to protein transport defects, protein misfolding, ER stress and markedly reducing the oxygen utilization of the mitochondria. Acetaldehyde promotes cell death by depleting the GSH reserve, inducing lipid peroxidation and increasing the level of free radicals.

Excess NADH causes a number of metabolic disorders including hyperlactacidemia, and hyperuricemia. NADH also opposes gluconeogenesis, leading to hypoglycemia, elevation of alpha-glycerophosphate level and inhibition of fatty acid oxidation. Inhibition of fatty acid oxidation favors steatosis and hyperlipidemia paving the way to the development of alcoholic liver disease (ALD). Acetaldehyde-protein adducts promote collagen production and contribute to fibrosis in the alcoholic liver.

Canonical ethanol metabolizing enzymes have been extensively studied, but our understanding of concomitant changes in oxido-reductive cellular milieu, epigenetic status, effects on global energy homeostasis, interactions with synergizing biological pathways, and their overall contribution to alcohol-induced diseases is lagging. Recently several lines of investigations provided convincing evidence that cellular changes accompanying ethanol metabolism may themselves regulate growth, development, and cellular homeostasis and induce organ pathologies. Notably, the enzymatic breakdown of ethanol affects the generation of several metabolites and cofactors that play seminal role in epigenetic gene regulation: NAD+, acetate, S-adenosylmethionine (SAM), and zinc. Fluctuation in the levels of these four critical compounds also causes energetic imbalance that may be a major contributor to alcohol induced organ pathologies. Additionally, the consequences of ethanol-induced gene expression changes due to reductive stress (decreased NAD+/NADH ratio), perturbation of 1-carbon metabolism (SAM), acetate and zinc utilization pathways in organ pathologies are largely unexplored.

A major goal of this FOA is to stimulate research that will broaden our understanding of how alcohol metabolism causes tissue damage, by considering not only the metabolic products of ethanol itself, but all of the biochemical changes resulting from alcohol metabolism in different cell types. The long-term goal is a comprehensive understanding of how alcohol metabolism impacts cellular function and end-organ disease, which has important implications for therapeutics development.

Ethanol and NAD+

Ethanol oxidation is accompanied by NAD+ reduction to NADH that generates highly reductive cytoplasmic environment in cells and tissues where ADH and CYP2E1 are active. This mechanism has been best studied in hepatocytes, but may be relevant in other cells as well. In mammals three major enzyme families are capable to cleave NAD+: sirtuins, ADP-ribose transferases including PARP, and cyclic ADP (cADP)-ribose synthase, two of which (sirtuins and PARPs) are also chromatin-modifying enzymes.

One major consequence of the decreased NAD+/NADH ratio is the inhibition of the NAD+ dependent SIRT1 histone deacetylase activity in mammalian cell, an orthologue of S. pombe Sir2 histone deacetylase, which links cellular metabolism status to transcriptional silencing. SIRT1 that requires NAD+ for its activity has essential functions in regulating circadian genes, caloric restriction and obesity pathways and consequently affecting longevity in every organism studied so far. Members of the mammalian sirtuin family of histone deacetylases (HDAC III) comprise seven homologues (SIRT1-7) with ubiquitous expression and diverse subcellular localization. Their functions affect a wide range of biological processes including tubulin deacetylation (SIRT2), adaptive thermogenesis, mitochondrial function, energy homeostasis, survival during genotoxic stress (SIRT3), genomic DNA stability and DNA repair (SIRT6). These considerations strongly suggest a role for SIRTs in alcohol-induced tissue injuries (e.g. Alcoholic Liver Disease, Fetal Alcohol Spectrum Disorders, ethanol-induced cancers) but their involvement has not been systematically investigated.

NAD+ dependent inhibition of SIRT HDACs increases global histone acetylation, as well as acetylation of key transcription factors and metabolic enzymes including SREBP-1c (Sterol Regulatory Binding Protein 1c), HSF1 (Heat Shock Factor 1) and Long-Chain Acyl Coenzyme A Dehydrogenase (LCAD). Collectively, these changes lead to metabolic disturbances and loss of protective capacity of the Heat Shock Response, contributing to ethanol-induced tissue injury. Therefore, understanding of organ specific histone code resulting from ethanol consumption, and the regulatory roles of protein acetylation are vital to successfully combat diverse alcoholic disorders.

Poly-ADP ribose polymerases (PARPs), which are the most abundant ADP-ribosyl transferases in mammalian cells, consist of a superfamily of up to 17 members. These enzymes are characterized by the highly conserved PARP domain that is the active site of the catalytic center. The best studied members are PARP 1 and 2: these ubiquitous zinc finger proteins are localized in the nucleus and are involved in chromatin maintenance and DNA repair by binding to single and double stranded breaks generated by genotoxic stress. Thus, PARPs may play a significant role in alcohol promoted cancers invoked by DNA damage of acetaldehyde. PARPs are the main modulators of intracellular NAD+ level, which in turn may indirectly control sirtuin functions through NAD+ availability. Changes in ADP-ribosylation of histone and other PARP target proteins brought upon by ethanol consumption are unknown and require further investigation. Furthermore, clarification of the functional interrelationship between SIRTs and PARPs mediated through NAD+ availability is needed. Conceivably, zinc deficiency, which is a well described consequence of excess ethanol ingestion, may affect PARP function, but this question has not been addressed experimentally.

Pioneering studies argue that excessive reductive environment itself can induce reductive stress and lead to potentially lethal cardiomyopathy in a transgenic mouse model and in human patients. This novel pathway may be operational in other tissues where reductive intracellular environment is generated by alcohol ingestion induced shift. The fundamental principles of this process are unclear and require further investigations.

Ethanol and acetate

As noted above, the two principal pathways of ethanol oxidation by ADH and CYP2E1 both yield highly reactive acetaldehyde that is further converted to acetate. Alcohol consumption can elevate acetate levels as much as 20-fold. Free acetate, the end product in the liver, is used then to generate acetyl-coenzyme A (acetyl-coA) catalyzed by cytosolic acetyl-coA synthase 1 (AceCS1) and mitochondrial acetyl-coA synthase 2 (AeCS2) in extrahepatic tissue. Acetyl-coA is ubiquitously utilized in histone acetylation, in the Krebs cycle, and fatty acid synthesis, as well as in post-translational acetylation of transcription factors, metabolic enzymes, cell motility factors, and other proteins. Acetylation and methylation are the best characterized posttranslational modifications of histone tails. Acetylation of various lysine residues of histones is associated with a transcriptionally permissive state by neutralizing the positive charge of histones and decreasing the interaction of the histone tails with the negatively charged phosphate groups of DNA. This, in turn leads to chromatin opening and increased accessibility of transcriptional machinery to chromatin template and exaggerated gene expression. Hyperacetylation of chromatin due to ethanol has been documented both in tissue culture cells and in different animal models of alcohol consumption, but the mechanism of hyperacetylation orchestrated by ethanol is largely unknown. Three mechanisms have been hypothesized: 1.) increased substrate availability (acetyl-CoA), 2.) modulation of histone acetyl transferases (HAT) and histone deacetylases (HDAC) 3.) increased enzymatic activity of AceCS1 and AceCS2 that may be operational. Most studies addressed the effect of alcohol at the global level and detected increased acetylation profiles, but studies analyzing specific genes affected by alcohol metabolism are just beginning to emerge. It is interesting to note that alcohol consumption seems to preferentially target certain histone lysine residues for acetylation, while others are unaffected. Understanding the mechanistic basis of this phenomenon is important for understanding how alcohol affects gene expression. Further investigations are needed to decipher the global alcohol and organ specific histone code under specific ethanol drinking paradigms. Regulatory effects of acetylation of numerous metabolic enzymes and transcription factors that are modified by alcohol drinking remain obscure and require further investigation.

Ethanol and SAM

SAM, the second most widely used co-factor in nature is synthesized in the liver from methionine and ATP by methionine adenosyltransferate (MAT) in the one-carbon metabolism pathway. SAM is the primary methyl donor in cells and is required for methylation of histones and other protein targets, DNA, RNA, biogenic amines, and phospholipids. This reaction, which can be carried out by a broad class of over a hundred methyltransferases, yields S-adenosylhomocysteine (SAH). Because SAH is a potent inhibitor of methyltransferases, it needs to be rapidly eliminated from the cells by SAH hydrolases. SAM plays fundamental role in epigenetic gene regulation by methylation of DNA, histones, enzymes, co-repressors, and co-activators of methylation pathways. Additionally, SAM is a precursor of glutathione, a small molecular weight endogenous antioxidant and a scavenger of ROS. Ethanol prominently alters the methione metabolism that is evidenced by changes in the intracellular concentration of SAM, SAH and other downstream metabolites. Alcohol induces SAM deficiency that has been associated with different stages of alcoholic liver injury. The mechanism of ALD due to SAM depletion is not fully understood, but GSH deficiency and increased ROS availability, activation of the Unfolded Protein Response (UPR) in the endoplasmic reticulum by hyperhomocystenemia, and hypomethylation of histones and DNA are hypothesized to contribute. SAM deficiency is mainly due to methionine adenosytransferase (MAT) and methionine synthase inhibition by ethanol that block the key steps in SAM biosynthesis that can be reversed by SAM or betaine feeding that restores the level of endogenous SAM. Although generally believed to be repressive, methylation of histone tails exhibits position specific effects at individual gene promoters: some are (e.g. H3K9) silencing, while others (e.g. H3K4) activating. How histon methylases are specifically recruited to a particular position on individual promoters to convey an ethanol signature is unclear and require further investigation. This notion also applies to the generation of alcohol induced hypomethylation signatures on promoter DNA.Compared to other post-translational modifications histone and DNA methylations are relatively irreversible epigenetic marks. Surprisingly, histone and DNA demethylases have only been recently identified and their reaction mechanisms appear to be quite complex and diverse. Among the three histone demethylases jumonji-domain containing (jmjC) demethylases are 2-oxoglutarate (2-OG) and Fe(II)-dependent dioxygenases, the lysine specific demethylase 1 (LSD1) is a flavine adenine dinucleotide (FAD)-dependent amine oxidase, while peptidyl arginine deiminase 4 (PAD4/PADI4) capacity as demethylase is still questioned.

An exciting development in recent years has been the identification of 5-hydroxy methyl cytosine (5-hmC) in mammalian DNA. The TET1 enzyme, product of an acute myeloid leukemia-associated gene, was recently identified as a 5-meC hydroxylase. Elegant studies demonstrated that TET1 is a 2-OG and Fe(II)-dependent dioxygenase that oxidizes 5-mC to 5-hmC, in a reaction similar to the jmjC histon demethylases. 5-hmC thus represents a new class of epigenetic modification with currently unknown function or may prompt DNA demethylation through an oxidative demethylation pathway. Since ethanol exposure modulates SAM and SAH level via changes in the NADH/NAD+ redox change, ethanol may also impact 5-hmC levels in different tissues as well. This mechanism and its possible role in alcohol-induced disease need further studies.

Ethanol and zinc

As an essential trace element, zinc plays a critical role in diverse biological processes including protein and carbohydrate metabolism, digestion, glucose control, immune system, wound healing, fertility, and growth. These effects are brought about by zinc’s modulation of gene expression, redox homeostasis and DNA repair, as it is a co-factor of about 300 different enzymes. Of particular importance, two groups of chromatin modifying enzymes require zinc for proper function: 1.) types I, II and IV histone deacetylases; and 2.) histone demethylases; both of them are inextricably associated with epigenetic gene regulation. Chronic and excessive ethanol intake is often related to zinc deficiency that is observed in ~50% of alcoholics due to decreased intestinal absorption, increased urinary excretion, and insufficient dietary zinc intake. Zinc deficiency is commonly observed in different stages of alcoholic liver disease (ALD). Recent studies clearly demonstrated a direct effect between zinc deficiency and histone acetylation level, but the molecular underpinnings of this process remain unclear. Both the recently identified LSD2 and jumonji-type histone demethylases also contain a zinc finger domain, which is hypothesized to be dramatically affected by zinc deficiency leading to pathological epigenetic landscape in chronic and heavy alcohol users. Zinc depletion may indirectly affect methyl group utilization by the inhibition of the zinc metalloenzyme betain-homocysteine S-methyl transferase that leads to general cellular hypomethylation. Further research is needed to extend our understanding how ethanol induced zinc deficiency contributes to the pathological gene programs and their phenotypic manifestation brought about by the inhibitory effects of alcohol on chromatin modifying enzymes.

Endogenous Substrates and Functions of Alcohol Metabolizing Pathways: Regulation of Growth, Development, and Disease

Mammals, including humans, have numerous highly elaborate and dynamic enzymatic pathways to break down ethanol and generate energy. Recently several lines of investigations provided convincing evidence that these enzymes may have additional functions beyond metabolizing exogenous alcohols, as they may have endogenous substrates that regulate cellular growth, development, and homeostasis. Ubiquitous, highly reactive endogenous aldehydes may induce DNA damage, a prelude to cancer, or perturb developmental processes leading to fetal damage e.g. fetal alcohol syndrome (FAS). Aldehyde dehydrogenase 2 (ALDH2) also provides cardioprotection. Consequently, genetic deficiencies in ALDH2 activities, and likely in other alcohol metabolizing enzymes, predispose the carriers to various genetic and developmental abnormalities including cardiovascular disease, bone marrow dysfunction, cancer and fetal alcohol syndrome. Understanding the molecular underpinnings of these cellular processes will provide opportunities to prevent or treat alcohol-induced organ damage and other alcohol related disorders.

Research Objectives

Research in the following key areas will help our understanding of the molecular underpinnings of these cellular processes and provide opportunities to prevent or treat alcohol-induced organ damage. Examples of research that might be supported under this FOA include, but are not limited to, the following:

The role of endogenous aldehydes and aldehyde dehydrogenases (ALDH) in DNA damage, cancer, and development (e.g. FAS)
The contribution and functional interplay of ALDH2 with DNA damage and cancer pathways
Mechanistic understanding the protective functions of ALDH2
Development of safe and effective ALDH2 activators (Alda) and inhibitors (Aldi) for treatment and/or prevention of alcohol induced pathologies
The effect of decreased NAD+/NADH ratio on NAD+ consuming enzymes: sirtuins, ADP-ribose transferases (PARP) and cyclic ADP (cADP)-ribose synthases and their effect of chromatin gene regulation
Develop sensitive and reliable in vivo assays to monitor NAD+ concentration in different cellular compartment
Determine how the cellular NAD+ pool is shared by NAD+ consuming enzymes
Investigation of reductive stress as a potential new paradigm in ethanol-related organ pathology
Regulation of SIRT1 targets by deacetylation: transcription factors, coactivators, and their effects on metabolic pathways by ethanol consumption (e.g., PGC-1alpha, FOXOs)
Does alcohol metabolism change Ac-CoA levels and increase histone acetylation as a consequence of acetate accumulation?
NAD+ and Ac-CoA energy sensor interplay in reprogramming of metabolic pathways
How ethanol alters the methionine pathway and leads to reduced SAM level and methylation in cells
Alcohol induced zinc deficiency and inhibition chromatin modifying enzymes
The effect of alcohol metabolism in NAD+/NADH ratio, and subsequent impact on sirtuins-related pathways in different organs and cell types
Understanding the mechanistic basis of how alcohol metabolism affects the epigenome, including 5-methyl cytosine and 5-hydroxy-methyl cytosine.
The effect of alcohol metabolism in various tissues at different stages of development, including in utero, fetal and embryonic development
Alcohol’s effect on redox state as well as its impact on metabolism of lipid, carbohydrate and overall energy balance
Suitable in vitro and in vivo experimental systems to model the diverse ways in which different cell types in the body are exposed to and metabolize ethanol
The role of genetic variation in the susceptibility to tissue injury, including carcinogenesis, resulting from alcohol metabolism
Understanding the effects of alcohol metabolism on the epigenome in different cells and organs, using either genome-wide or hypothesis-driven candidate gene approach
Mechanisms by which alcohol metabolism contributes to carcinogenesis in various organs
Role of central and peripheral circadian rhythms on alcohol-metabolism
Bioinformatics, computational, and mathematical modeling to understand and integrate experimental information on alcohol metabolism
Development of more effective biomarkers of alcohol metabolism
Alcohol metabolism as a therapeutic target
Understanding how alcohol metabolism in the liver affects the function of other organs in the body
Alcohol metabolism by the oral and colonic microbiome, as well as the effects of alcohol and its metabolites on the composition and function of the microbiome

Section II. Award Information

Funding Instrument	Grant: A support mechanism providing money, property, or both to an eligible entity to carry out an approved project or activity.
Application Types Allowed	New Renewal Resubmission Revision The OER Glossary and the SF424 (R&R) Application Guide provide details on these application types.
Funds Available and Anticipated Number of Awards	The number of awards is contingent upon NIH appropriations and the submission of a sufficient number of meritorious applications.
Award Budget	Application budgets are not limited but need to reflect the actual needs of the proposed project.
Award Project Period	The maximum period is 5 years.

NIH grants policies as described in the NIH Grants Policy Statement will apply to the applications submitted and awards made in response to this FOA.

Section III. Eligibility Information

1. Eligible Applicants

Eligible Organizations

Higher Education Institutions

Public/State Controlled Institutions of Higher Education
Private Institutions of Higher Education

The following types of Higher Education Institutions are always encouraged to apply for NIH support as Public or Private Institutions of Higher Education:

Hispanic-serving Institutions
Historically Black Colleges and Universities (HBCUs)
Tribally Controlled Colleges and Universities (TCCUs)
Alaska Native and Native Hawaiian Serving Institutions
Asian American Native American Pacific Islander Serving Institutions (AANAPISIs)

Nonprofits Other Than Institutions of Higher Education

Nonprofits with 501(c)(3) IRS Status (Other than Institutions of Higher Education)
Nonprofits without 501(c)(3) IRS Status (Other than Institutions of Higher Education)

For-Profit Organizations

Small Businesses
For-Profit Organizations (Other than Small Businesses)

Governments

State Governments
County Governments
City or Township Governments
Special District Governments
Indian/Native American Tribal Governments (Federally Recognized)
Indian/Native American Tribal Governments (Other than Federally Recognized)
Eligible Agencies of the Federal Government
U.S. Territory or Possession

Other

Independent School Districts
Public Housing Authorities/Indian Housing Authorities
Native American Tribal Organizations (other than Federally recognized tribal governments)
Faith-based or Community-based Organizations
Regional Organizations
Non-domestic (non-U.S.) Entities (Foreign Institutions)

Foreign Institutions

Non-domestic (non-U.S.) Entities (Foreign Institutions) are eligible to apply.
Non-domestic (non-U.S.) components of U.S. Organizations are eligible to apply.
Foreign components, as defined in the NIH Grants Policy Statement, are allowed.

Required Registrations

Applicant Organizations

Applicant organizations must complete and maintain the following registrations as described in the SF 424 (R&R) Application Guide to be eligible to apply for or receive an award. All registrations must be completed prior to the application being submitted. Registration can take 6 weeks or more, so applicants should begin the registration process as soon as possible. The NIH Policy on Late Submission of Grant Applications states that failure to complete registrations in advance of a due date is not a valid reason for a late submission.

Dun and Bradstreet Universal Numbering System (DUNS) - All registrations require that applicants be issued a DUNS number. After obtaining a DUNS number, applicants can begin both SAM and eRA Commons registrations. The same DUNS number must be used for all registrations, as well as on the grant application.
System for Award Management (SAM) (formerly CCR) Applicants must complete and maintain an active registration, which requires renewal at least annually. The renewal process may require as much time as the initial registration. SAM registration includes the assignment of a Commercial and Government Entity (CAGE) Code for domestic organizations which have not already been assigned a CAGE Code.
NATO Commercial and Government Entity (NCAGE) Code Foreign organizations must obtain an NCAGE code (in lieu of a CAGE code) in order to register in SAM.
eRA Commons - Applicants must have an active DUNS number and SAM registration in order to complete the eRA Commons registration. Organizations can register with the eRA Commons as they are working through their SAM or Grants.gov registration. eRA Commons requires organizations to identify at least one Signing Official (SO) and at least one Program Director/Principal Investigator (PD/PI) account in order to submit an application.
Grants.gov Applicants must have an active DUNS number and SAM registration in order to complete the Grants.gov registration.

Program Directors/Principal Investigators (PD(s)/PI(s))

All PD(s)/PI(s) must have an eRA Commons account. PD(s)/PI(s) should work with their organizational officials to either create a new account or to affiliate their existing account with the applicant organization in eRA Commons. If the PD/PI is also the organizational Signing Official, they must have two distinct eRA Commons accounts, one for each role. Obtaining an eRA Commons account can take up to 2 weeks.

Eligible Individuals (Program Director/Principal Investigator)

Any individual(s) with the skills, knowledge, and resources necessary to carry out the proposed research as the Program Director(s)/Principal Investigator(s) (PD(s)/PI(s)) is invited to work with his/her organization 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 support.

For institutions/organizations proposing multiple PDs/PIs, visit the Multiple Program Director/Principal Investigator Policy and submission details in the Senior/Key Person Profile (Expanded) Component of the SF424 (R&R) Application Guide.

2. Cost Sharing

This FOA does not require cost sharing as defined in the NIH Grants Policy Statement.

3. Additional Information on Eligibility

Number of Applications

Applicant organizations may submit more than one application, provided that each application is scientifically distinct.

NIH will not accept any application that is essentially the same as one already reviewed within the past thirty-seven months (as described in the NIH Grants Policy Statement), except for submission:

To an RFA of an application that was submitted previously as an investigator-initiated application but not paid;
Of an investigator-initiated application that was originally submitted to an RFA but not paid; or
Of an application with a changed grant activity code.

Section IV. Application and Submission Information

1. Requesting an Application Package

Applicants must download the SF424 (R&R) application package associated with this funding opportunity using the Apply for Grant Electronically button in this FOA or following the directions provided at Grants.gov.

2. Content and Form of Application Submission

It is critical that applicants follow the instructions in the SF424 (R&R) Application Guide, except where instructed in this funding opportunity announcement to do otherwise. Conformance to the requirements in the Application Guide is required and strictly enforced. Applications that are out of compliance with these instructions may be delayed or not accepted for review.

For information on Application Submission and Receipt, visit Frequently Asked Questions Application Guide, Electronic Submission of Grant Applications.

Page Limitations

All page limitations described in the SF424 Application Guide and the Table of Page Limits must be followed

Required and Optional Components

The forms package associated with this FOA includes all applicable components, required and optional. Please note that some components marked optional in the application package are required for submission of applications for this FOA. Follow all instructions in the SF424 (R&R) Application Guide to ensure you complete all appropriate optional components.

Instructions for Application Submission

The following section supplements the instructions found in the SF424 (R&R) Application Guide and should be used for preparing an application to this FOA.

SF424(R&R) Cover