U01AA029965
Cooperative Agreement
Overview
Grant Description
6/11 Astrocyte-Specific Changes and Interventions in Alcohol Dependence - Project Summary
Astrocytes respond to CNS damage and disease with changes in gene expression and morphology and with immune activation to become reactive astrocytes. Alcohol use disorder is characterized in part by neuroimmune responses that increase with the progression of the disorder. Reactive astrocytes upregulate the expression of tissue-type plasminogen activator (TPA) encoded by the gene PLAT, which is involved in brain plasticity, the remodeling of the brain extracellular matrix, and neuroimmune responses including microglial activation and neuroinflammation. TPA is upregulated by ethanol in several brain areas and in astrocytes in several models of ethanol exposure.
The main scientific questions driving the proposed studies are:
1) What are the changes in translating RNA occurring in astrocytes after chronic intermittent ethanol-2 bottle choice (CIE-2BC)?
2) What are the changes in nuclear gene expression occurring in astrocytes after CIE-2BC?
3) Does PLAT/TPA knock-down in astrocytes reduce escalation in drinking, neuroimmune responses, and synaptic changes induced by CIE-2BC?
We hypothesize that CIE-2BC induces changes in the translation of neuroimmune genes in astrocytes and that some, but not all, changes in translation are driven by changes in transcription. We also hypothesize that knocking down PLAT selectively in astrocytes will attenuate escalation in drinking, induction of neuroimmune genes, and synaptic changes induced by CIE-2BC.
We propose to use the ALDH1L1-EGFP-RPL10A mouse model that allows the selective pull down of actively translating RNA from astrocytes by the translating ribosome affinity purification (TRAP) method. Moreover, this mouse line has GFP fluorescence in the nucleus that allows for the isolation of astrocyte-specific nuclei by fluorescent-activated cell sorting (FACS).
In Specific Aim 1, we will study the translatome in amygdala and PFC of female and male ALDH1L1-EGFP-RPL10A mice after CIE-2BC by TRAP-RNA-Seq. Pathway analysis will be performed to determine enrichment in the biological processes affected by CIE-2BC. We will employ TRAP-QPCR, Western blot, and immunohistochemistry (IHC) to validate changes in neuroimmune gene translation and protein expression.
In Specific Aim 2, we will study the nuclear transcriptome in the amygdala and PFC of female and male ALDH1L1-EGFP-RPL10A mice after CIE-2BC by FACS sorting of astrocyte nuclei followed by RNA-Seq. We will integrate transcriptome and translatome data to identify RNAs regulated by alcohol through transcription-dependent and transcription-independent mechanisms. We will employ FACS-QPCR and fluorescence in situ hybridization (FISH)-RNAscope to validate changes in neuroimmune gene expression.
In Specific Aim 3, we will investigate the hypothesis that TPA knock-down in astrocytes attenuates CIE-2BC-induced escalation of drinking, immune response, and synaptic changes. Inducible ALDH1L1-Cre/ERT2 mice will be crossed to floxed PLAT mice for the selective knock down of PLAT in astrocytes. Mice lacking TPA in astrocytes will undergo CIE-2BC; drinking escalation, immune response, and synaptic changes will be assessed.
Astrocytes respond to CNS damage and disease with changes in gene expression and morphology and with immune activation to become reactive astrocytes. Alcohol use disorder is characterized in part by neuroimmune responses that increase with the progression of the disorder. Reactive astrocytes upregulate the expression of tissue-type plasminogen activator (TPA) encoded by the gene PLAT, which is involved in brain plasticity, the remodeling of the brain extracellular matrix, and neuroimmune responses including microglial activation and neuroinflammation. TPA is upregulated by ethanol in several brain areas and in astrocytes in several models of ethanol exposure.
The main scientific questions driving the proposed studies are:
1) What are the changes in translating RNA occurring in astrocytes after chronic intermittent ethanol-2 bottle choice (CIE-2BC)?
2) What are the changes in nuclear gene expression occurring in astrocytes after CIE-2BC?
3) Does PLAT/TPA knock-down in astrocytes reduce escalation in drinking, neuroimmune responses, and synaptic changes induced by CIE-2BC?
We hypothesize that CIE-2BC induces changes in the translation of neuroimmune genes in astrocytes and that some, but not all, changes in translation are driven by changes in transcription. We also hypothesize that knocking down PLAT selectively in astrocytes will attenuate escalation in drinking, induction of neuroimmune genes, and synaptic changes induced by CIE-2BC.
We propose to use the ALDH1L1-EGFP-RPL10A mouse model that allows the selective pull down of actively translating RNA from astrocytes by the translating ribosome affinity purification (TRAP) method. Moreover, this mouse line has GFP fluorescence in the nucleus that allows for the isolation of astrocyte-specific nuclei by fluorescent-activated cell sorting (FACS).
In Specific Aim 1, we will study the translatome in amygdala and PFC of female and male ALDH1L1-EGFP-RPL10A mice after CIE-2BC by TRAP-RNA-Seq. Pathway analysis will be performed to determine enrichment in the biological processes affected by CIE-2BC. We will employ TRAP-QPCR, Western blot, and immunohistochemistry (IHC) to validate changes in neuroimmune gene translation and protein expression.
In Specific Aim 2, we will study the nuclear transcriptome in the amygdala and PFC of female and male ALDH1L1-EGFP-RPL10A mice after CIE-2BC by FACS sorting of astrocyte nuclei followed by RNA-Seq. We will integrate transcriptome and translatome data to identify RNAs regulated by alcohol through transcription-dependent and transcription-independent mechanisms. We will employ FACS-QPCR and fluorescence in situ hybridization (FISH)-RNAscope to validate changes in neuroimmune gene expression.
In Specific Aim 3, we will investigate the hypothesis that TPA knock-down in astrocytes attenuates CIE-2BC-induced escalation of drinking, immune response, and synaptic changes. Inducible ALDH1L1-Cre/ERT2 mice will be crossed to floxed PLAT mice for the selective knock down of PLAT in astrocytes. Mice lacking TPA in astrocytes will undergo CIE-2BC; drinking escalation, immune response, and synaptic changes will be assessed.
Funding Goals
TO DEVELOP A SOUND FUNDAMENTAL KNOWLEDGE BASE WHICH CAN BE APPLIED TO THE DEVELOPMENT OF IMPROVED METHODS OF TREATMENT AND MORE EFFECTIVE STRATEGIES FOR PREVENTING ALCOHOLISM AND ALCOHOL-RELATED PROBLEMS. THE NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM (NIAAA) SUPPORTS RESEARCH IN A BROAD RANGE OF DISCIPLINES AND SUBJECT AREAS RELATED TO BIOMEDICAL AND GENETIC FACTORS, PSYCHOLOGICAL AND ENVIRONMENTAL FACTORS, ALCOHOL-RELATED PROBLEMS AND MEDICAL DISORDERS, HEALTH SERVICES RESEARCH, AND PREVENTION AND TREATMENT RESEARCH. SMALL BUSINESS INNOVATION RESEARCH (SBIR) PROGRAM: TO INCREASE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, TO INCREASE SMALL BUSINESS PARTICIPATION IN FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION. SMALL BUSINESS TECHNOLOGY TRANSFER (STTR) PROGRAM: TO STIMULATE AND FOSTER SCIENTIFIC AND TECHNOLOGICAL INNOVATION AND TECHNOLOGY TRANSFER THROUGH COOPERATIVE RESEARCH AND DEVELOPMENT CARRIED OUT BETWEEN SMALL BUSINESS CONCERNS AND RESEARCH INSTITUTIONS, TO INCREASE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION.
Grant Program (CFDA)
Awarding / Funding Agency
Place of Performance
Oregon
United States
Geographic Scope
State-Wide
Related Opportunity
Analysis Notes
Amendment Since initial award the total obligations have increased 300% from $377,845 to $1,511,380.
Oregon Health & Science University was awarded
6/11 Astrocyte-specific changes and interventions in alcohol dependence
Cooperative Agreement U01AA029965
worth $1,511,380
from National Institute on Alcohol Abuse and Alcoholism in March 2022 with work to be completed primarily in Oregon United States.
The grant
has a duration of 4 years 10 months and
was awarded through assistance program 93.273 Alcohol Research Programs.
The Cooperative Agreement was awarded through grant opportunity Integrative Neuroscience Initiative on Alcoholism (INIA) Consortia (U01 Clinical Trial Optional).
Status
(Ongoing)
Last Modified 8/20/25
Period of Performance
3/15/22
Start Date
1/31/27
End Date
Funding Split
$1.5M
Federal Obligation
$0.0
Non-Federal Obligation
$1.5M
Total Obligated
Activity Timeline
Transaction History
Modifications to U01AA029965
Additional Detail
Award ID FAIN
U01AA029965
SAI Number
U01AA029965-1076216703
Award ID URI
SAI UNAVAILABLE
Awardee Classifications
Public/State Controlled Institution Of Higher Education
Awarding Office
75N500 NIH National Institute on Alcohol Abuse and Alcoholism
Funding Office
75N500 NIH National Institute on Alcohol Abuse and Alcoholism
Awardee UEI
NPSNT86JKN51
Awardee CAGE
0YUJ3
Performance District
OR-90
Senators
Jeff Merkley
Ron Wyden
Ron Wyden
Budget Funding
| Federal Account | Budget Subfunction | Object Class | Total | Percentage |
|---|---|---|---|---|
| National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Health and Human Services (075-0894) | Health research and training | Grants, subsidies, and contributions (41.0) | $755,690 | 100% |
Modified: 8/20/25