Identifying new targets for the treatment of alcohol dependence and relapse: epigenetic analysis of the abstinent brain

确定治疗酒精依赖和复发的新靶点：戒酒大脑的表观遗传学分析

• 批准号: 10553449
• 负责人: Rita P Cervera Juanes
• 金额: $ 42.55万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553449
• 关键词: Abstinence; Advanced Development; Affect; Alcohol abuse; Alcohol consumption; Alcohol withdrawal syndrome; Alcoholic beverage heavy drinker; Alcohols; Attention; Behavioral; Brain; Cause of Death; Chronic; DNA Methylation; Data; Development; Disease remission; Drug Metabolic Detoxication; Electrophysiology (science); Epigenetic Process; Ethanol; Exposure to; FDA approved; Female; Gene Expression; Gene Proteins; Gene Targeting; Genes; Glutamates; Goals; Heavy Drinking; Intoxication; Lead; Link; Macaca; Maps; Measures; Mediating; Modeling; Molecular; Mus; Nature; Nucleus Accumbens; Outpatients; Pathway interactions; Persons; Pharmaceutical Preparations; Pharmacology; Population; Primates; Progressive Disease; Proteins; Publishing; Relapse; Reporting; Rodent; Rodent Model; Sampling; Self Administration; Sequence Analysis; Signal Transduction; Support Groups; Synaptic plasticity; Testing; Time; Tissues; Transcript; Translations; United States; Up-Regulation; Variant; alcohol abuse therapy; alcohol exposure; alcohol relapse; alcohol use disorder; bisulfite; brain tissue; chronic alcohol ingestion; dependence relapse; design; differential expression; drinking; drinking behavior; effective therapy; gamma-Aminobutyric Acid; genetic manipulation; genome-wide; male; negative emotional state; neural circuit; neuroadaptation; neurotransmission; new therapeutic target; novel; patch clamp; preference; prevent; preventable death; relapse risk; sex; success; synaptic function; tool; transcriptome sequencing; transcriptomics; transmission process

SUMMARY Alcohol use disorder (AUD) affects more than 12% of the US population and is the fourth leading preventable cause of death in the US. AUD is associated with compulsive drinking and emergence of a negative emotional state during abstinence, resulting in two-thirds of afflicted people relapsing within months of completing alcohol cessation treatment. Repeated cycles of intoxication and abstinence are linked to persistent alterations in the brain that remain long after detoxification, which drive relapse of drinking, often at levels higher than before abstinence. Identifying the molecular mechanisms that lead to hazardous alcohol use and contribute to the high rates of relapse is imperative for the effective design of better treatments to curb alcohol abuse. Our previous studies illustrate the power of integrating epigenetic, transcriptomic, circuitry, pharmacology and behavioral tools to elucidate the molecular underpinnings contributing to hazardous alcohol use. Those studies, using genome- wide DNA methylation (DNAm) analysis of the nucleus accumbens (NAc) to compare macaque alcohol-naive and alcohol-drinkers, identified differential DNAm (D-DNAm) signals, mapping to genes not previously linked to alcohol use, but with high relevance to synaptic plasticity modulation. Experimental manipulation of two of these DNAm-linked genes resulted in altered glutamate and GABA neurotransmission and changes in ethanol intake. The the next critical step is to understand how repeated abstinence/relapse cycles alters DNAm signals and how these changes contribute to adaptations in neurocircuitry. We hypothesize that some D-DNAm signals associated with chronic alcohol use may persist during abstinence, contributing to risk of relapse. In addition, de novo D-DNAm signals generated during repeated cycles of abstinence/relapse, may further heighten relapse risk. To identify abstinence-associated DNAm, we will use the highly translational macaque alcohol self- administration model and our proven genome-wide approach to identify D-DNAm in the NAc of macaques following > 12 months of chronic alcohol self-administration and then 3 cycles of forced abstinence (each cycle: 1 month abstinence and 3 months of open-access). After integrating gene and transcript variant expression and D-DNAm data generated from the same subjects and tissues, a subset of compelling, novel targets will be selected for functional study using pharmacological and genetic manipulation approaches in a mouse chronic intermittent ethanol (CIE) model. This rodent CIE model has been extensively used to model cycles of alcohol exposure and withdrawal leading to an escalation of drinking during relapse. In addition to recording ethanol intake, the effects of target manipulation on neurotransmission will be evaluated using patch-clamp electrophysiological analysis. Overall, these studies will aid in our understanding of the molecular mechanism(s) establishing risk for relapse. This information will be critical to advancing the development of effective therapies for alcohol abuse and to prevent relapse.

概括 酒精使用障碍 (AUD) 影响超过 12% 的美国人口，是第四大可预防疾病 美国的死因。 AUD 与强迫性饮酒和负面情绪的出现有关 戒酒期间的状态，导致三分之二的患者在戒酒后几个月内复发 戒烟治疗。反复的中毒和戒酒循环与体内的持续改变有关。 解毒后大脑中的残留物仍存在很长时间，这会导致饮酒复发，并且其水平通常比以前更高 节制。确定导致危险酒精使用和导致高浓度酒精的分子机制 复发率对于有效设计更好的治疗方法来遏制酗酒至关重要。我们之前的 研究表明整合表观遗传学、转录组、电路、药理学和行为工具的力量 阐明导致有害酒精使用的分子基础。这些研究使用基因组- 对伏隔核 (NAc) 进行广泛 DNA 甲基化 (DNAm) 分析，以比较未接触酒精的猕猴 和饮酒者，鉴定出差异 DNAm (D-DNAm) 信号，映射到先前未关联的基因 饮酒，但与突触可塑性调节高度相关。对其中两个的实验操作 DNAm 相关基因导致谷氨酸和 GABA 神经传递的改变以及乙醇摄入量的变化。 下一个关键步骤是了解重复的戒断/复发周期如何改变 DNAm 信号以及如何 这些变化有助于神经回路的适应。我们假设一些 D-DNAm 信号 与长期饮酒相关的疾病可能在戒酒期间持续存在，从而导致复发的风险。此外，德 在重复的戒断/复发周期中产生的 novo D-DNAm 信号可能会进一步加剧复发 风险。为了识别与禁欲相关的 DNAm，我们将使用高度翻译的猕猴酒精自体 管理模型和我们经过验证的全基因组方法来识别猕猴 NAc 中的 D-DNAm 经过> 12个月的长期自我饮酒，然后进行3个周期的强制戒酒（每个周期： 1 个月禁欲和 3 个月开放获取）。整合基因和转录变体表达并 从相同受试者和组织生成的 D-DNAm 数据，引人注目的新颖目标的子集将被 选择使用药理学和遗传操作方法对小鼠慢性病进行功能研究 间歇乙醇（CIE）模型。这种啮齿动物 CIE 模型已广泛用于模拟酒精循环 暴露和戒断导致复发期间饮酒量增加。除了记录乙醇 摄入后，将使用膜片钳评估目标操纵对神经传递的影响 电生理分析。总的来说，这些研究将有助于我们理解分子机制 确定复发风险。这些信息对于推进有效疗法的开发至关重要 防止酗酒并防止复发。