Epigenetic Mechanisms of Negative Affective State of AUD

AUD负性情感状态的表观遗传机制

基本信息

批准号：
10613977
负责人：
SUBHASH C. PANDEY
金额：
$ 19.43万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10613977
关键词：
ATAC-seq Acetylation Alcohol consumption Alcohol dependence Alcohol withdrawal syndrome Amygdaloid structure Anatomy Antibodies Anxiety Attenuated Autopsy Behavior Biological Assay Biological Process CREBBP gene Cell Differentiation process Cell Nucleus ChIP-seq Chemicals Chromatin Chromatin Structure Chronic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Convulsions Coupled DNA DNA Damage DNA Methylation DUSP6 protein Data Data Set Dendritic Spines Development EP300 gene Epigenetic Process Equilibrium Ethanol Ethanol dependence Female Funding G9a histone methyltransferase Gene Expression Gene Expression Regulation Genes Genetic HDAC4 gene HMGB1 gene Histone Acetylation Histone Deacetylase Inhibitor Histones Human Infusion procedures Lead Lysine Maintenance Measures Medial Messenger RNA Methylation Modification Molecular Molecular Target Neuroimmune Neurons Pathway Analysis Pathway interactions Pharmacotherapy Play Progress Reports Proteins Rattus Regional Anatomy Regulation Relapse Research Risk Risk Factors Rodent Role Self Administration Small Interfering RNA Synapses Synaptic plasticity Techniques Testing Tissue-Specific Gene Expression Translating Transposase Tremor Withdrawal Withdrawal Symptom Withholding Treatment alcohol exposure alcohol research alcohol use disorder anoctamin 5 anxiety-like behavior associated symptom attenuation chronic alcohol ingestion designer receptors exclusively activated by designer drugs drinking drinking behavior epigenetic regulation epigenome experimental study gene function gene network genome-wide genomic locus histone acetyltransferase histone deacetylase 2 histone modification male negative affect next generation sequencing novel pre-clinical preclinical study premature prevent problem drinker transcriptome transcriptome sequencing transcriptomics translational study

项目摘要

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.

项目摘要：新兴的临床前和临床证据表明与酒精有关的症状提取/负面影响是复发和维持酒精使用障碍（AUD）的主要危险因素。解剖结构包括扩展的杏仁核，尤其是杏仁核的中央核（CEA），与焦虑和饮酒行为有着强烈的影响。表观遗传机制，例如组蛋白和 DNA化学修饰已被证明在基因表达的调节中起重要作用。这护理的研究成分将研究如何由慢性酒精诱导的表观遗传修饰在全基因组水平上的暴露和退出导致调节基因网络通路异常杏仁核中的各种生物学过程，产生焦虑般的行为并升级酒精喝。我们提出以下具体目的：1）检查a）染色质可及性的状态和使用ATAC-SEQ和H3K27ME3芯片seq的基因组表观遗传标记（男性和女性）慢性乙醇暴露后乙醇提取期间。新兴数据集将与现有的 RNA-Seq和ChIP-Seq（H3K9/14AC标记）以鉴定差异基因的综合表观遗传调节在杏仁核中的表达。 b）双重特异性磷酸酶6（DUSP6，通过合并确定的新基因 H3K9/14AC芯片seq/rna-seq）siRNA将通过激活CBP来减轻焦虑的行为并增加乙醇抽出期间杏仁核基因的组蛋白乙酰化。 2）检查a）g9a siRNA输注在CEA中，将减弱类似焦虑的行为，使表观遗传和基因表达变化以及在乙醇抽出期间，大鼠杏仁核中突触和树突状刺的缺陷； b）神经元使用化学发生（GQ耦合Dreadd）方法在CEA中刺激会减弱焦虑样通过恢复HDAC2/CBP调节中适当平衡的行为，从而导致组蛋白增加乙醇撤离期间杏仁核中靶基因的乙酰化和c）指导神经元特异性P300HAT 通过CRISPR-DCAS9输注CEA将增加靶基因的组蛋白乙酰化，从而导致基因增加在杏仁核中的表达，焦虑样行为的衰减以及戒断期间的饮酒升级。 3）检查CEA输注HDAC2和G9A siRNA是否会减弱乙醇依赖性诱导的通过手术乙醇自身给药衡量的雄性和雌性大鼠饮酒的升级。 4）到将酒精依赖大鼠杏仁核中基因的表观遗传动力学和表达转化为尸体人类酒精中毒的杏仁核与饮酒数据相关。拟议研究的完成将提供有关杏仁核整个转录组表观遗传调节的新信息，导致鉴定出AUD药物疗法的分子靶标。