PA-14-138: Neuroimmune Mechanisms of Alcohol Related Disorders (R21)

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	Neuroimmune Mechanisms of Alcohol Related Disorders (R21)
Activity Code	R21 Exploratory/Developmental Research Grant
Announcement Type	Reissue of PA-11-065
Related Notices	May 10, 2017 - New NIH "FORMS-E" Grant Application Forms and Instructions Coming for Due Dates On or After January 25, 2018. See NOT-OD-17-062. NOT-OD-16-004 - NIH & AHRQ Announce Upcoming Changes to Policies, Instructions and Forms for 2016 Grant Applications (November 18, 2015) NOT-OD-16-006 - Simplification of the Vertebrate Animals Section of NIH Grant Applications and Contract Proposals (November 18, 2015) NOT-OD-16-011 - Implementing Rigor and Transparency in NIH & AHRQ Research Grant Applications (November 18, 2015) 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-14-138
Companion Funding Opportunity	PA-14-139, R01 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	This Funding Opportunity Announcement (FOA) encourages Exploratory/Developmental Research Project Grant (R21) applications from institutions/organizations that propose to study the neuroimmune mechanisms of alcohol related disorders. Studies using animal models and post-mortem human alcoholic brains suggest that alcohol exposure alters the neuroimmune system in the brain. However, it remains unclear how the altered neuroimmune signaling contributes to brain functional and behavioral changes associated with alcohol dependence. Recent studies reveal that neuroimmune molecules are expressed in neurons and glia, and play an important role in modulating synaptic function, neurodevelopment, and neuroendocrine function. These neuromodulatory properties, together with their essential roles in neuroinflammation, provide a new frame work to understand the role of neuroimmune factors in mediating neuroadaptation and behavioral phenotypes associated with alcohol use disorders. Studies supported by this FOA will provide fundamental insights of neuroimmune mechanisms underlying brain functional and behavioral changes induced by alcohol.

Key Dates

Posted Date	March 3, 2014
Open Date (Earliest Submission Date)	May 16, 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	May 8, 2017
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

Purpose

This Funding Opportunity Announcement (FOA) encourages grant applications to study neuroimmune mechanisms underlying alcohol-related disorders. Emerging data indicate that the function of neuroimmune factors goes beyond their established role in neuroinflammation and that they also play an important role in various brain functions. It is now known that neuroimmune factors are expressed in neurons and glia, and play a critical role in modulating synaptic function, neurodevelopment, and neuroendocrine function. These neuromodulatory properties, together with their essential role in neuroinflammation, provide a new frame work to understand neuroimmune mechanisms underlying brain functional and behavioral changes induced by alcohol exposure. This FOA seeks studies to address the role of neuroimmune modulation in response to and in the neuroadaptation of the brain to acute and chronic alcohol exposure, and alcohol dependence and withdrawal.

The R21 mechanism is intended to encourage new exploratory and developmental research projects. For example, such projects could assess the feasibility of a novel area of investigation or a new experimental system that has the potential to enhance health-related research. Another example could include the unique and innovative use of an existing methodology to explore a new scientific area. These studies may involve considerable risk but may lead to a breakthrough in a particular area, or to the development of novel techniques, agents, methodologies, models, or applications that could have a major impact on a field of biomedical, behavioral, or clinical research. An R21 grant application need not have extensive background material or preliminary information.

Background

Rapid advances in understanding neuroimmune function have far reaching impact on research in many areas of neuroscience. It is becoming increasingly clear that neuroimmune factors modulate a wide range of brain functions and play an important role in development, normal brain function, and CNS dysfunctions, including neurodegenerative diseases, neuropsychiatric disorders, and drug abuse. Studies using human alcoholic brains, gene knockout mice, and gene expression profiling have revealed a clear link between alcohol exposure and altered neuroimmune function. However, it is essentially unknown how neuroimmune changes contribute to brain functional changes due to alcohol exposure. In addition to their primary role in mediating neuroinflammation, neuroimmune factors, such as cytokines, chemokines, and MHC, are critical for a variety of normal brain functions. They are expressed in neurons and glia and regulate synaptic function, mediate neuron-glia communication, interact with neuroendocrine and neuropeptide systems, and regulate neurogenesis and CNS development.

Neuroimmune Modulation of Synaptic Function

Many immune molecules, including chemokines, cytokines, and type I major histocompatibility complex (MHC), interact with neurotransmitter systems and play essential roles in modulating synaptic function. As a family of G-protein coupled receptor systems, binding of chemokines to chemokine receptors triggers a cascade of signaling events, which subsequently modulate neurotransmitter release and activities of many receptors and channels. For example, chemokines CCL2 and CXCL12 regulate the release of several neurotransmitters, including glutamate, GABA, and dopamine. The chemokine receptor CCR2 cross-desensitizes GABAa and mu-opioid receptors. In addition, activation of CXCR4 by its ligand CXCL12 triggers the release of glutamate from astrocytes. Studies also demonstrated that proinflamatory cytokines regulate synaptic transmission and plasticity, and contribute to the maintenance of the homeostasis of neuronal networks. For example, TNFα differentially modulates trafficking of AMPA type glutamate receptors and GABA receptors. It facilitates excitatory synaptic transmission, while decreasing inhibitory synaptic transmission. Blocking of IL-1 with IL-1 receptor antagonist prevents the formation of long-term potentiation. In addition, as the key component of the neuroimmune system, microglia dynamically detect the brain environment, even at the resting state, and contribute to postnatal development, neuroplasticity, and circuit function. It becomes clear that the bi-directional communication between neuron and microglia plays important roles in both normal brain function and neurobiological diseases. Thus, neuroimmune system modulates synaptic function by presynaptic, postsynaptic, and neural glial mechanisms. Such actions of neuroimmune system offer potential neuroimmune mechanisms for brain functional changes due to excessive alcohol exposure.

Neuroimmune Factors and Neuroinflammation

Neuroimmune factors have significant impact on neuroinflammation. They exert either neuroprotective or neurotoxic effect via distinct mechanisms. Upon insult by environmental toxins or neuronal damage, microglia can be at different stages of activation. At the initial stage of the innate immune response, TNFα and IL1 are two main cytokines that are produced by glia. They exert neuroprotective effects by promoting the maturation of oligodendrocytes and increasing the secretion of neurotrophines. However, over-activated microglia release numerous proinflammatory cytokines, chemokines, and inducible nitric oxide synthase, which synergistically mediate neuroinflammation. To counterbalance the neuroinflammation, the brain produces anti-inflammatory factors, such as IL-10 and transforming growth factor-b1, to inhibit inflammatory responses. It is important to note that neuroinflammation may not only be provoked by pathological conditions but also be trigged by increased neuronal activities, such as those associated with noxious stimuli, psychological stress and epileptic seizures.

Neuroimmune Molecules in Neurodevelopment

Neuroimmune molecules are involved in all stages of neurodevelopment. They are expressed in both the developing and adult brain and play critical roles in neuro and glio genesis, neuronal migration, and axonal path finding. Dysregulation of CNS immune molecules at the early stage of brain development cause significant behavioral deficits, which are evident by the increased risk of several neurological disorders. Cytokines and chemokines play diverse roles in embryonic brain development and adult neurogenesis. For example, gp130 family cytokines and TNFα regulate neurogenesis, gliogenesis, and neuronal survival in the embryonic brain. Chemokine CXCL-12 is considered an indispensable chemoattractant for neuronal migration and axonal path finding in the developing nervous system. Disruption of certain chemokine receptors causes the malformation of granule cell layers of the cerebellum, the dentate gyrus, and cortical interneurons. In the adult brain, cytokine TNFα and IL-6 inhibit neurogenesis, whereas the constitutive expression of IL-1 is critical for hippocampal neurogenesis. In addition, the chemokine CXCL-12 and its receptor CXCR4 are expressed in the subventricular zone and regulate migration and proliferation of progenitor cells. Evidence suggests that alcohol exposure disrupts cytokine profile during early neuronal differentiation and influences adult neurogenesis, and alters the neuroimmune gene expression in a brain regional dependent manner. The essential role of neuroimmune molecules in neurodevelopment and adult neurogenesis provide potential mechanisms to understand the effects of alcohol on CNS development.

Neuroimmune Factors Modulate Neuroendocrine Function

The involvement of neuroimmune molecules in the regulation of neuroendocrine function is demonstrated in animal models and by human conditions of stress and depression. Increased levels of cytokines are associated with depression and sickness behavior. The underlying molecular and cellular mechanisms of these conditions are believed to be primarily due to the dysregulation of the HPA axis, as well as serotonergic and dopaminergic systems, by neuroimmune factors. A variety of cytokines have potent effects on the HPA axis by regulating the release of neuropeptides and neurohormones, including corticotrophin release factor, ACTH, or cortisol. Conversely, glucocorticoids induced by chronic stress have a significant impact on the neuroimmune system by regulating expressions of cytokines in the hippocampus, the prefrontal cortex, and the hypothalamus. In addition, several chemokines are expressed in the paraventricular nucleus of the hypothalamus and regulate the stress-related neuroendocrine responses, such as the release of arginine vasopressin. One hallmark of alcohol dependence is the dysregulated HPA axis with altered release of CRF, vasopressin, ACTH and cortisol due to ethanol exposure. The ability of neuroimmune molecules in regulating the HPA axis suggests that neuroimmune molecules may play an integrative role in the close link between stress responses and alcohol dependence.

Impact of Ethanol Exposure on the Neuroimmune System

Studies using animal models and post-mortem human alcoholic brains suggest that alcohol exposure has a significant impact on the neuroimmune system in the brain. Expressions of several immune related genes are altered in human alcoholic brains and are differentially correlated with the high and low alcohol consuming rodent lines. In addition, polymorphisms of genes encoding IL-1 and IL-1 receptor antagonist, as well as the gene for an anti-inflammatory cytokine IL-10, are associated with the susceptibility to alcoholism. Furthermore, interruption of certain neuroimmune gene expression reduced alcohol consumption in rodents. These studies provide a clear link between altered neuroimmune networks and alcoholism. However, it remains unclear how these genetic variations are linked to the high alcohol intake and how alcohol exposure causes adaptive changes of neuroimmune networks in the brain.

Recent studies provide strong evidence that ethanol directly targets the CNS neuroimmune networks. For example, ethanol activates TLR4 dependent signaling pathways in microglia and triggers the production of a variety of neuroimmune factors. A study using a mouse binge drinking model revealed a long lasting increase of the chemokine CCL2 and the cytokine TNFα but a decrease of the anti-inflammatory cytokine IL-10 in the mouse brain. In addition, chronic alcohol exposure induces neuroimmune activation through HMGB1/TLR signaling. Together, these studies provide molecular and cellular evidence that ethanol alters the neuroimmune system in the brain. Moreover, in vivo animal studies provide further evidence that neuroimmune modulation contributes to alcohol dependence. Recent studies demonstrated that cytokines facilitate alcohol withdrawal-induced anxiety via the CRF signaling in the central amygdala and that pharmacological suppressions of various neuroimmune signaling pathways reduce alcohol consumption.

In summary, alcohol exposure has significant effects on the neuroimmune system. Neuroimmune activation may contribute to alcohol dependence via a variety of mechanisms. The neuromodulatory properties of neuroimmune factors, together with their primary roles in mediating neuroinflammation, provide a new frame work to understand the role of neuroimmune factors in mediating brain functional changes contributing to excessive alcohol drinking and alcoholism.

Research Objectives

Research areas of interest to NIAAA include, but are not limited to:

Molecular, cellular and network

Determine the effects of acute, chronic alcohol exposure, alcohol dependence or withdrawal on neuroimmune system at the molecular, cellular and functional level.
Determine the effects of alcohol on neuroimmune factor-regulated neurotransmitter release, synaptic function, and communication among neurons, glia, and progenitor cells.
Examine genetic and epigenetic mechanisms underlying neuroimmune dysregulation associated with alcohol dependence.
Examine the impact of alcohol exposure on the interaction between the neuroimmune and neuroendocrine systems, e.g., hypothalamic gene expression, HPA activity, neuropeptide and neurohormone release, and glucocorticoid mediated signaling.
Define the impact of alcohol-induced neuroimmune changes on structural and functional connectivity at the cellular and circuitry level.
Examine the cross-talk between the neuroimmune system and neurotransmitter or neuromodulatory systems known to mediate alcohol dependence.
Reveal mechanisms underlying the distinct impact of alcohol on the CNS neuroimmune system and the peripheral immune system.

Neurodevelopment

Examine the role of neuroimmune factors in the developing and adult brain, e.g. neurogenesis, synaptic pruning, neurocircuit formation, axon guidance, and neuronal migration in the context of alcohol exposure.
Determine the long-term consequences of early alcohol-induced neuroimmue alteration on brain development and function.

Behavior and other neurological conditions

Determine the role of neuroimmune modulation in binge drinking, alcohol dependence, withdrawal, and relapse.
Examine the role of neuroimmune signaling in stress-induced alcohol relapse and alcohol-withdrawal-induced anxiety.
Determine the contributions of alcohol-induced neuroimmune changes to neurological disorders, e.g., cognitive impairment, anxiety, depression, and PTSD.
Examine the involvement of the neuroimmune system in alcohol-induced sleep disorders.

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 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. RFA with only one IC:
Award Budget	The combined budget for direct costs for the two year project period may not exceed $275,000. No more than $200,000 may be requested in any single year. Applicants may request direct costs in $25,000 modules, up to the total direct costs limitation of $275,000 for the combined two-year award period.
Award Project Period	The award project period may not exceed 2 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