Distinguishing preexistent and induced epigenetic risk for alcohol use disorders

区分酒精使用障碍的先存风险和诱发的表观遗传风险

• 批准号: 10553537
• 负责人: Rita P Cervera Juanes
• 金额: $ 58.05万
• 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: 2022-01-22 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10553537
• 关键词: Accounting; Address; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohols; Area; Autopsy; Behavior; Behavioral; Biopsy; Brain; Cause of Death; Cells; Chronic; Classification; Cytosine; DNA Methylation; Data; Dependence; Detection; Development; Dose; Electrophysiology (science); Epigenetic Process; Female; Future; Gene Expression; Gene Expression Profile; Gene Proteins; Gene Targeting; Genes; Goals; Hair; Heavy Drinking; Heterogeneity; Human; Individual; Interdisciplinary Study; Lead; Link; Macaca; Macaca mulatta; Measures; Microfluidics; Modeling; Modification; Molecular; Mus; Neurologic; Neurotransmitters; Nucleus Accumbens; Persons; Pharmacological Treatment; Pharmacology; Prefrontal Cortex; Prevention strategy; Primates; RNA; Recovery; Research; Resources; Risk; Rodent Model; Role; Sampling; Self Administration; Sequence Analysis; Signal Transduction; Time; Tissue Sample; Tissues; Transcript; United States; Viral Vector; Work; alcohol abuse therapy; alcohol availability; alcohol consequences; alcohol effect; alcohol risk; alcohol use disorder; base; behavioral study; bisulfite; bisulfite sequencing; brain tissue; design; differential expression; drinking; effective therapy; genome-wide; hazardous drinking; insight; male; mouse model; neuroadaptation; neuromechanism; new therapeutic target; non-alcoholic; nonhuman primate; novel; patch clamp; problem drinker; relating to nervous system; success; targeted treatment; transcriptome sequencing

SUMMARY Nearly 88,000 people die from alcohol-related causes annually, making it the fourth leading preventable cause of death in the United States. Understanding the molecular mechanisms underlying the neurological changes that lead to alcohol dependence is crucial for developing new, effective alcohol use disorder (AUD) treatments. Towards this goal, recent studies have identified genome-wide DNA methylation (DNAm) signals distinguishing alcoholic and non-alcoholic post-mortem brain tissue. In addition, a subset of the differential DNAm (D-DNAm) signals was associated with the altered expression of nearby genes, suggesting that DNAm regulates the gene expression patterns associated with alcoholic dependence. However, it is unknown whether some of the DNAm signals detected in the post-mortem tissue were preexistent, potentially contributing risk for hazardous drinking, rather than being induced by chronic alcohol. This study aims to distinguish these two possibilities, and to pinpoint the functional consequences of alcohol-induced DNAm in the prefrontal cortex (PFC). The study makes use of the highly relevant and well-characterized nonhuman primate (NHP) alcohol self- administration model. In this model, voluntary alcohol consumption levels are measured in rhesus macaques daily for ~12 months, enabling the accurate classification of natural “low” and “heavy” alcohol drinkers. In addition, a small portion of the PFC is biopsied prior to alcohol access, providing opportunity to establish baseline measures of DNAm. Genome-wide bisulfite sequencing (GWBS) will be used to identify alcohol-dose dependent, differentially methylated cytosines (DMCs) and regions (DMRs) in the PFC of the rhesus macaques. The same approach will be used to compare DNAm in PFC obtained from the macaques prior to alcohol access, enabling the detection of preexistent, alcohol-dose associated DNAm signatures. In addition, the macaque D-DNAm signals will be compared to D-DNAm data generated from post-mortem human alcoholic PFC tissue, identifying alcohol-associated DNAm that is replicated across two primate species. Next, the expression of DMC/DMR-associated genes, including both whole gene and alternative transcript expression, will be correlated with the DNAm data. Based on support from the human-macaque DNAm comparison, and corresponding expression data, a subset of compelling, novel gene targets will be selected for functional study. Target gene activity will be modulated using pharmacological and gene expression modification approaches in a murine model. The neural effects of target modulation will be evaluated using patch-clamp electrophysiological analysis, and alcohol use will be evaluated using intermittent alcohol, 2 bottle choice models. In total, this work will identify preexistent and alcohol-induced DNAm modifications in the primate PFC and will clarify the role of a subset of DNAm-linked genes in promoting alcohol use. The findings of this study will provide functional support for the design of promising new AUD treatments.

概括 每年有近 88,000 人死于与酒精相关的原因，使其成为第四大可预防原因 了解美国死亡的分子机制。 导致酒精依赖的因素对于开发新的、有效的酒精使用障碍（AUD）治疗方法至关重要。 为了实现这一目标，最近的研究已经确定了全基因组 DNA 甲基化 (DNAm) 信号，可区分 此外，酒精和非酒精死后脑组织还存在差异 DNAm (D-DNAm) 的子集。 信号与附近基因的表达相关，表明 DNAm 调节该基因 然而，尚不清楚其中一些是否与酒精依赖相关。 在死后组织中检测到的 DNAm 信号是预先存在的，可能会造成危险 饮酒，而不是由长期饮酒引起的这项研究旨在区分这两种可能性， 并查明酒精诱导的 DNAm 对前额皮质 (PFC) 的功能影响。 研究利用高度相关且特征明确的非人类灵长类动物 (NHP) 酒精自我 在此模型中，测量了恒河猴的自愿饮酒水平。 每天大约 12 个月，能够准确分类自然“低度”和“重度”饮酒者。 此外，在获取酒精之前对一小部分 PFC 进行活检，从而提供了确定 DNAm 基线测量（GWBS）将用于确定酒精剂量。 恒河猴 PFC 中依赖的差异甲基化胞嘧啶 (DMC) 和区域 (DMR) 相同的方法将用于比较之前从猕猴获得的 PFC 中的 DNAm。 酒精访问，能够检测预先存在的、与酒精剂量相关的 DNAm 特征。 猕猴 D-DNAm 信号将与死后人类生成的 D-DNAm 数据进行比较 酒精性 PFC 组织，识别出在两种灵长类动物中复制的酒精相关 DNAm。 DMC/DMR 相关基因的表达，包括全基因和替代转录本 表达，将基于人类-猕猴 DNAm 的支持与 DNAm 数据相关联。 比较和相应的表达数据，将选择引人注目的新颖基因目标的子集 用于功能研究。将使用药理学和基因表达来调节靶基因活性。 将使用小鼠模型中的修改方法来评估目标调节的神经效应。 膜片钳电生理分析，并使用间歇性酒精（2 瓶）评估酒精使用情况 总的来说，这项工作将识别 DNAm 中预先存在的和酒精诱导的修饰。 灵长类动物 PFC 并将阐明 DNAm 相关基因子集在促进饮酒中的作用。 这项研究将为设计有前景的新型 AUD 治疗方法提供功能支持。