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，使其成为第四个主要可预防原因 在美国死亡。了解神经系统变化的分子机制 导致酒精依赖性对于发展新的，有效的酒精使用障碍（AUD）治疗至关重要。 为了实现这一目标，最近的研究确定了全基因组DNA甲基化（DNAM）的信号 酗酒和非酒精后尸体后脑组织。另外，差分DNAM的子集（d-dnam） 信号与附近基因的表达改变有关，表明DNAM调节基因 与酒精依赖相关的表达模式。但是，尚不清楚某些 验尸组织中检测到的DNAM信号既有存在 喝酒，而不是由慢性酒精诱发。这项研究旨在区分这两种可能性： 并查明酒精诱导的DNAM在前额叶皮层（PFC）中的功能后果。 研究利用了高度相关和表征良好的非人类灵长类动物（NHP）酒精自我 管理模型。在此模型中，自愿性饮酒水平以恒河猕猴进行测量 每天持续约12个月，可以准确地分类自然的“低”和“重”酒精。 此外，PFC的一小部分是在酒精获取前进行活检的，提供了建立的机会 DNAM的基线度量。全基因组的亚硫酸盐测序（GWBS）将用于鉴定酒精剂量 恒河猴PFC中的依赖性，不同的甲基化胞嘧啶（DMC）和区域（DMR） 猕猴。将使用相同的方法比较从猕猴中获得的DNAM 酒精获取，可以检测前期酒精剂量相关的DNAM特征。此外， 将猕猴D-DNAM信号与验尸人类产生的DNAM数据进行比较 醇PFC组织，鉴定出在两个主要物种中复制的酒精相关的DNAM。下一个， DMC/DMR相关基因的表达，包括整个基因和替代转录本 表达式将与DNAM数据相关。基于人类马克克dnam的支持 比较和相应的表达数据，将选择引人注目的新基因靶标的子集 用于功能研究。靶基因活性将使用药物和基因表达进行调节 鼠模型中的修改方法。目标调节的神经效应将使用 贴片钳电生理分析和饮酒将使用间歇性酒精，2瓶进行评估 选择模型。总的来说，这项工作将确定在 灵长类动物PFC并将阐明DNAM-C-C-C-C-C-C-C-C-C-C-C-C-C-C-C-Chink基因在促进饮酒中的作用。发现 这项研究将为有希望的新AUD治疗的设计提供功能支持。