Thursday, April 25, 2024
4/25/2024
ABSTRACT Several clinical and preclinical studies suggest that adolescent binge drinking is one of the major risk factors for the development of psychiatric disorders, including anxiety and alcohol use disorder (AUD) later in life. Adolescence represents an important stage of brain development, and epigenetic mechanisms are known to orchestrate developmental maturation of the brain from adolescence to adulthood. This research component of Neurobiology of Adolescent Drinking in Adulthood (NADIA) consortium will examine how epigenetic modifications induced by adolescent intermittent ethanol (AIE) at the genome-wide level lead to an aberrant gene network pathway regulating various synaptic mechanisms in the amygdala that are responsible for anxiety- like and alcohol drinking behavior in adulthood. This component will identify closed and open domains of the epigenome after AIE in adult amygdala using Assay for Transposase-Accessible Chromatin-sequencing (ATAC- seq), and the corresponding regulation of the transcriptome by merging this dataset with existing RNA-seq data. The overarching hypothesis of this proposal is that AIE will produce differential gene expression due to altered status of the epigenome in adult amygdala. The dynamic changes in epigenetic targets (HATs/EZH2/G9a/LSD1/DNMT3b) will induce changes in the transcriptome that regulate synaptic mechanisms in the amygdala after AIE in adulthood, ultimately regulating anxiety-like and alcohol- drinking behaviors. Specific Aim 1 will examine a) the status of chromatin accessibility and loci of genomic epigenetic marks using ATAC-seq in the adult amygdala of rats (male & female) after AIE. The emerging data set will be merged with existing RNA-seq data to identify epigenetically regulated transcriptomic changes, and b) To validate expression and epigenetic enrichments of new genes in the amygdala after AIE and use them for functional studies. Specific Aim 2 will examine the effects of neuronal epigenomic editing in the CeA using CRISPR/dCas9-p300, CRISPR/dCas9-KRAB, CRISPR/dCas9-Lsd1 (histone acetylation/methylation), or CRISPR/dCAS9-Tet1 (DNA methylation mechanism) on AIE-induced changes in gene expression and on anxiety-like and alcohol drinking behaviors in adult male and female rats. Specific Aim 3 will examine whether treatment with G9a inhibitor (UNC0642) will normalize epigenetic, gene expression, and dendritic spine changes in the amygdala and attenuate AIE-induced anxiety-like and alcohol drinking behaviors in adult male and female rats. Finally, Specific Aim 4 will translate epigenetic dynamics and expression of novel genes that will be identified from RNA-seq and ATAC-seq in the AIE rat amygdala to post-mortem amygdala of human alcoholics with early age onset and correlate them to drinking data (daily or weekly ethanol intake). The proposed studies will provide new information about epigenetic regulation of the whole transcriptome in the amygdala after AIE, leading to identification of epigenetic targets for drug development for the treatment of adult psychopathology.
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