# Epigenetic Mechanisms of Negative Affective State of AUD

> **NIH NIH P50** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2021 · $194,325

## Abstract

Project Summary:
The emerging preclinical and clinical evidence suggests that symptoms associated with alcohol
withdrawal/negative affect is a primary risk factor for relapse and maintenance of alcohol use disorder (AUD).
Anatomical structures comprising the extended amygdala, particularly the central nucleus of amygdala (CeA),
are strongly implicated in anxiety and alcohol-drinking behaviors. Epigenetic mechanisms such as histone and
DNA chemical modifications, have been shown to play important roles in the regulation of gene expression. This
research component of CARE will examine how epigenetic modifications induced by chronic alcohol
exposure and withdrawal at genome-wide level lead to an aberrant gene network pathway regulating
various biological processes in the amygdala, producing anxiety-like behavior and escalated alcohol
drinking. We propose the following Specific Aims: 1) To examine a) status of chromatin accessibility and loci of
genomic epigenetic marks using ATAC-seq and H3K27me3 ChIP-seq in the amygdala of rats (male & female)
during ethanol withdrawal after chronic ethanol exposure. The emerging data set will be merged with existing
RNA-seq and ChIP-seq (H3K9/14ac mark) to identify integrated epigenetic regulation of differential gene
expression in the amygdala. b) Dual specificity phosphatase 6 (Dusp6, novel gene identified by merger of
H3K9/14ac ChIP-seq/RNA-seq) siRNA will attenuate anxiety-like behaviors via activating CBP and increasing
histone acetylation of genes in the amygdala during ethanol withdrawal. 2) To examine if a) G9a siRNA infusion
in the CeA will attenuate anxiety-like behaviors, normalize epigenetic and gene expression changes as well as
deficits in synapses and dendritic spines in the amygdala of rats during ethanol withdrawal; b) Neuronal
stimulation in the CeA using a chemogenetic (Gq coupled DREADD) approach will attenuate anxiety-like
behaviors via restoring the proper balance in HDAC2/CBP regulation thereby leading to increased histone
acetylation of target genes in the amygdala during ethanol withdrawal and c) Neuron-specific p300HAT guided
by CRISPR-dCas9 infusion into CeA will increase histone acetylation at target genes, leading to increased gene
expression in the amygdala, attenuation of anxiety-like behaviors and escalated drinking during withdrawal. 3)
To examine whether CeA infusion of a HDAC2 and G9a siRNA will attenuate ethanol dependence-induced
escalation in drinking in male and female rats, as measured by operant ethanol-self administration. 4) To
translate epigenetic dynamics and expression of genes in the amygdala of alcohol dependent rats to postmortem
amygdala of human alcoholics and correlate them to drinking data. Completion of proposed studies will
provide new information on the epigenetic regulation of the whole transcriptome in the amygdala,
leading to identification of molecular targets for the development of pharmacotherapy of AUD.

## Key facts

- **NIH application ID:** 10147640
- **Project number:** 5P50AA022538-07
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** SUBHASH C. PANDEY
- **Activity code:** P50 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $194,325
- **Award type:** 5
- **Project period:** 2015-04-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10147640

## Citation

> US National Institutes of Health, RePORTER application 10147640, Epigenetic Mechanisms of Negative Affective State of AUD (5P50AA022538-07). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10147640. Licensed CC0.

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