Epigenetic Mechanisms of Positive Affective State of AUD

AUD积极情感状态的表观遗传机制

• 批准号: 10613969
• 负责人: MARK S BRODIE
• 金额: $ 19.81万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 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/10613969
• 关键词: ATAC-seq; Abstinence; Acetylation; Affective; Alcohol abuse; Alcohol consumption; Alcohol withdrawal syndrome; Amygdaloid structure; Anxiety; Area; Autopsy; Behavioral; Biological; Brain; CRISPR/Cas technology; ChIP-seq; Cholesterol; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Coupled; DNA Methylation; Data; EP300 gene; EZH2 gene; Electrophysiology (science); Enzymes; Epigenetic Process; Ethanol; Exhibits; Funding; Gene Cluster; Gene Expression; Gene Expression Regulation; Genes; Genetic; Goals; HDAC2 gene; Hippocampus; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone-Lysine N-Methyltransferase; Histones; Human; Intervention; Lead; Link; Methods; Methylation; Modification; Molecular; Motivation; Neurons; Nucleus Accumbens; Pathway Analysis; Pathway interactions; Phenotype; Physiology; Play; Prefrontal Cortex; Proteins; Publishing; Rattus; Recording of previous events; Relapse; Rewards; Role; Sampling; Small Interfering RNA; Substance Withdrawal Syndrome; Symptoms; Synapses; Technology; Testing; Translating; Ventral Tegmental Area; Withdrawal; Work; alcohol craving; alcohol exposure; alcohol research; alcohol seeking behavior; alcohol use disorder; cell type; chromatin immunoprecipitation; clinically relevant; designer receptors exclusively activated by designer drugs; drinking; drinking behavior; effective therapy; enzyme pathway; epigenetic regulation; epigenome; experimental study; gamma-Aminobutyric Acid; gene network; genetic technology; genome sequencing; histone methylation; methyl group; neurosteroids; novel; pharmacologic; pre-clinical; preclinical study; prevent; problem drinker; promoter; transcriptome; transcriptome sequencing; whole genome

Project Summary As a hallmark of alcohol use disorder (AUD), the withdrawal syndrome includes both negative symptoms like anxiety and motivational symptoms such as craving for alcohol, which promotes relapse and alcohol seeking. Understanding how alcohol withdrawal changes brain physiology is an important step towards developing effective treatments to reduce alcohol abuse. The ventral tegmental area (VTA) is an important brain area that projects to components of the extended amygdala, including the nucleus accumbens, prefrontal cortex, amygdala and hippocampus. Changes in the physiology of VTA neurons induced by withdrawal may underlie the alcohol-seeking during AUD. The sensitivity of neurons of the VTA to inhibition by gamma aminobutyric acid (GABA) is decreased during alcohol withdrawal but normalized by histone deacetylase (HDAC) inhibitors, indicating epigenetic changes induced by withdrawal in the VTA may be amenable to pharmacological manipulation. Whole genome sequencing and molecular studies of this project identified a cluster of genes that encode cholesterol synthesis pathway enzymes that showed decreased expression during alcohol withdrawal, and are functionally relevant to decreased GABA sensitivity in VTA neurons during withdrawal. The proposed project plans to further characterize the role of specific epigenetic modifications in withdrawal-induced regulation of genes responsible for cholesterol synthesis, and the effect of these changes in withdrawal-induced GABA hyposensitivity and drinking behaviors. By using electrophysiological, behavioral, and state of the art molecular biological methods such as chemogenetics and CRISPR technology, we anticipate achieving the following goals: 1) to reveal novel epigenetic marks and changes in gene expression associated with chronic alcohol exposure and withdrawal with whole genome approaches, 2) to examine whether decreased histone acetylation and increased histone methylation reduce expression of cholesterol synthesis enzymes in the VTA during withdrawal in a cell-type specific manner, and probe the role of cholesterol synthesis enzyme genes in withdrawal-related drinking behavior and GABA hyposensitivity, 3) to determine whether targeted epigenetic intervention counteracts withdrawal-induced phenotypes by using CRISPR to prevent decreases in histone acetylation on promoters of key cholesterol synthesis enzyme genes, and 4) to translate to post-mortem human alcoholic VTA the epigenetic dynamics and expression of genes related to GABA hyposensitivity that have been identified in the rat VTA. Ultimately, these studies are needed to understand epigenetic adaptation of VTA neurons involved in the positive affective state during alcohol withdrawal, and, with the other components of this Center, will provide a great deal of information on epigenetic mechanisms involved in withdrawal-induced brain changes, and possible pharmacological approaches toward more effective treatment of AUD.

项目摘要 作为酒精使用障碍的标志（AUD），戒断综合征包括两个负面症状，例如 焦虑和动机症状，例如渴望酒精，这会促进复发和寻求酒精。 了解饮酒如何改变脑生理是发展的重要一步 有效的治疗方法来减少酗酒。腹侧盖区（VTA）是一个重要的大脑区域 向扩展杏仁核的组成部分的项目，包括伏隔核，前额叶皮层， 杏仁核和海马。戒断引起的VTA神经元生理学的变化可能是 澳元期间寻求酒精。 VTA神经元对γ氨基丁酸抑制的敏感性 （GABA）在戒酒期间降低，但通过组蛋白脱乙酰基酶（HDAC）抑制剂标准化， 表明VTA戒断引起的表观遗传变化可能与药理学有关 操纵。该项目的整个基因组测序和分子研究确定了一组基因 编码胆固醇合成途径酶，在戒酒期间表达降低， 并且在戒断期间VTA神经元中的GABA灵敏度降低在功能上与之相关。提议 项目计划，以进一步表征特定表观遗传修饰在提款诱导的调节中的作用 负责胆固醇合成的基因，以及这些变化在戒断诱导的GABA中的影响 低敏和饮酒行为。通过使用电生理，行为和艺术分子状态 化学遗传学和CRISPR技术等生物学方法，我们预计实现以下目标： 1）揭示与慢性酒精暴露有关的新型表观遗传标记和基因表达的变化 并使用整个基因组方法提取，2）检查组蛋白乙酰化降低和 增加组蛋白甲基化减少了戒断期间VTA中胆固醇合成酶的表达 以细胞类型的特异性方式，并探测胆固醇合成酶基因在戒断相关的作用 饮酒行为和GABA低敏，3）确定是否有针对性的表观遗传干预 通过使用CRISPR来防止组蛋白乙酰化在 关键胆固醇合成酶基因的启动子，4）转化为验尸后的人类酒精VTA 在 大鼠VTA。最终，需要这些研究来了解VTA神经元的表观遗传适应 在戒酒期间的积极情感状态下，与该中心的其他组成部分，将 提供大量有关戒断引起的大脑变化的表观遗传机制的信息， 以及更有效治疗AUD的可能的药理方法。