Center for Alcohol Research in Epigenetics

Project Summary: Alcohol use disorder (AUD) can be characterized by a pattern of compulsive alcohol drinking and development of several withdrawal symptoms that promote relapse and maintenance of addiction. Long-term alcohol use causes molecular, structural, and functional changes in key brain regions such as medial prefrontal cortex (mPFC), hippocampus, amygdala, and ventral tegmental area (VTA) which may drive behavioral phenotypes, such as anxiety, depression, panic, pain, deficits in cognition and reduced reward sensitivity. Several studies have shown that protein families such as histone deacetylases (HDACs), histone acetyltransferases (HATs), histone methyltransferases (HMTs), DNA methyltransferases (DNMTs), and lysine demethylases are important players in epigenetic and transcriptional changes during withdrawal after chronic ethanol exposure. However, the cell- type specificity of the interplay between epigenetic mechanisms (open chromatin accessibility) and the transcriptomic changes that occur across brain regions [prelimbic mPFC, central nucleus of amygdala (CeA), dorsal hippocampus, and VTA] which eventually leads to the development and maintenance of AUD is not well known. The overall aim of this Alcohol Research Center is to evaluate the cellular heterogeneity of transcriptomic and epigenomic changes leading to structural and electrophysiological changes in specific brain regions that underlie withdrawal behavioral phenotypes in AUD. This renewal P50 application entitled “Center for Alcohol Research in Epigenetics (CARE)” consists of four highly inter-related preclinical and translational research projects [research project #1, VTA, (Brodie /Grayson), research project #2, Amygdala, (Pandey/Sparta), research project #3, Hippocampus, (McMurray/ Krishnan) and research project #4, mPFC, (Glover/Auta)] and two Cores [Administrative (Pandey/ Brodie), and Epigenetics Core (Grayson/Maienschein- Cline/Gao/Boergens/Krishnan)] utilizing animal models of AUD and human postmortem brain regions of AUD subjects. In addition, the CARE will continue to serve as an important resource and provide a scientifically enriched environment for training opportunities for the next generation of alcohol researchers and will disseminate scientific knowledge of AUD to the general public through a community outreach program. The primary thematic focus of CARE as a whole will be to causally link emerging cell type-specific mechanisms identified by 10x genomics (snRNA-seq and snATAC-seq) in the proposed brain circuitry to several behavioral phenotypes of AUD during withdrawal. Mechanistic approaches (CRISPR-dCas9 or shRNA) will be used to manipulate the cell type-specific epigenome through alterations of either histone acetylation/methylation mechanisms or DNA methylation mechanisms, in key brain circuitry that regulates behavioral phenotypes associated with AUD (anxiety, depression, panic, pain, cognition deficits and reduced reward sensitivity) in order to better understand the pathophysiology of AUD and develop better pharmacotherapy.