Identifying Brain Epitranscriptomic Changes Associated with Alcohol Use Disorder

识别与酒精使用障碍相关的大脑表观转录组变化

• 批准号: 10580861
• 负责人: Huiping Zhang
• 金额: $ 56.17万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580861
• 关键词: Adult; Affect; Air; Alcohol consumption; Alcohols; American; Amygdaloid structure; Animal Model; Autopsy; Brain; Brain region; Caucasians; Cell physiology; Cells; Cerebellum; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA Methylation; Dependence; Digestion; Disease; Engineering; Environmental Risk Factor; Epigenetic Process; Ethanol; Female; Gene Expression; Genes; Genetic Transcription; Genetic Variation; Goals; Guide RNA; Hippocampus; Human; Immune response; Individual; Messenger RNA; Methylation; MicroRNAs; Modeling; Modification; Morbidity - disease rate; Morphology; Mus; Neurons; Nucleotides; Nucleus Accumbens; Pathway interactions; Patients; Pharmacological Treatment; Pilot Projects; Prefrontal Cortex; RNA; RNA Splicing; RNA methylation; Regulation; Research; Research Design; Resolution; Rewards; Role; Sampling; Self Administration; Signal Transduction; Site; Stimulus; Therapeutic; Tissue Sample; Translating; Translations; Ventral Tegmental Area; Vulnerable Populations; addiction; alcohol effect; alcohol exposure; alcohol risk; alcohol use disorder; base; brain tissue; chronic alcohol ingestion; demethylation; design; disorder risk; epitranscriptome; epitranscriptomics; genetic risk factor; histone modification; innovation; insight; male; methylome; methylomics; mortality; mouse model; novel; nuclease; pharmacologic; posttranscriptional; putamen; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY/ABSTRACT Alcohol use disorder (AUD) affects about 4.2% (or 14.1 million) of American adults each year and causes substantial morbidity and mortality. While genetic variation can influence an individual's vulnerability to AUD, chronic alcohol consumption can also lead to alcohol tolerance and dependence, but the underlying mechanism is unclear. There is evidence that that chronic alcohol use alters DNA methylation of specific genes, leading to gene expression changes and possibly an increased risk of AUD. RNA methylation is a common post- transcriptional modification, and it responds rapidly to a variety of stimuli and translates stimulatory signals into cellular activity. We hypothesize that alcohol use alters an individual's RNA methylome (or epitranscriptome), resulting in altered expression of genes involved in AUD-related pathways. The objective of this research is to explore messenger RNA (mRNA) methylomic changes in the brain of AUD subjects and analyze the effect of mRNA methylation on mRNA expression and neuronal activity, thus providing evidence for a new gene expression regulatory mechanism of AUD. To achieve this goal, we propose three specific aims. First, we will profile mRNA methylomic and transcriptomic changes in eight regions (amygdala, caudate, cerebellum, hippocampus, nucleus accumbens, prefrontal cortex, putamen, and ventral tegmental area) of postmortem brains of AUD subjects. Second, we will use chronic intermittent ethanol (CIE)-exposed mice as models to verify AUD-associated brain mRNA methylation and expression changes as well as the correlation of ethanol exposure-induced mRNA methylation and expression changes with the escalation of ethanol self-administration in mice. Third, we will employ a novel epitranscriptome editing approach to investigate the effect of AUD- associated mRNA methylation changes on mRNA expression and neuronal activity. We expect to (1) observe AUD-associated mRNA methylation and expression changes in specific (or across) brain regions; (2) verify AUD- associated mRNA methylation and expression changes by mouse modeling; and (3) confirm the functional role of AUD-associated mRNA methylation in regulating mRNA expression and neuronal activity. Our study design is innovative. AUD-associated mRNA methylation changes will be examined at single-base resolution. An integrative study approach (human postmortem brain studies, mouse modeling, and epitranscriptome editing) will be employed. The proposed research will provide evidence about the influence of brain epitranscriptomic changes on AUD risk. Given that mRNA methylation is dynamic and reversible, mRNAs with differential methylation in the brain of AUD subjects could be potential targets for AUD treatment by pharmacologically altering mRNA methylation status.

项目总结/摘要 酒精使用障碍（AUD）每年影响约4.2%（或1410万）的美国成年人， 发病率和死亡率很高。虽然遗传变异可以影响个体对AUD的脆弱性， 长期饮酒也会导致酒精耐受和依赖，但其潜在机制 还不清楚有证据表明，长期饮酒会改变特定基因的DNA甲基化，导致 基因表达改变，可能增加AUD的风险。RNA甲基化是一种常见的 转录修饰，它迅速响应各种刺激，并将刺激信号翻译成 细胞活动我们假设酒精的使用改变了个体的RNA甲基化组（或表转录组）， 导致AUD相关通路中涉及的基因表达改变。本研究的目的是 探索AUD受试者脑中信使RNA（mRNA）甲基化组学变化，并分析 mRNA甲基化对mRNA表达和神经元活性的影响，从而为新基因的发现提供了证据 AUD的表达调控机制。为了实现这一目标，我们提出了三个具体目标。一是 在八个区域（杏仁核，尾状核，小脑， 海马、前额叶皮质、壳核和腹侧被盖区 AUD受试者的大脑。其次，我们将使用慢性间歇性乙醇（CIE）暴露小鼠作为模型来验证 AUD相关脑mRNA甲基化和表达变化以及乙醇的相关性 乙醇诱导的mRNA甲基化和表达随乙醇自身给药的增加而变化 对小鼠第三，我们将采用一种新的epitranscriptome编辑方法来研究AUD的作用， 相关的mRNA甲基化改变对mRNA表达和神经元活性的影响。我们希望（1）观察 特定（或跨）脑区域中AUD相关mRNA甲基化和表达变化;（2）验证AUD- 通过小鼠模型相关mRNA甲基化和表达变化;和（3）确认功能作用 AUD相关mRNA甲基化在调节mRNA表达和神经元活性中的作用。我们的研究设计 是创新的。将以单碱基分辨率检查AUD相关mRNA甲基化变化。一个 综合研究方法（人类死后大脑研究，小鼠建模和epitranscriptome编辑） 将被雇用。这项研究将提供证据，说明脑epitranscriptomic 澳元风险的变化。考虑到mRNA甲基化是动态和可逆的，具有差异的mRNA甲基化可能是不可逆的。 AUD受试者大脑中的甲基化可能是通过EMT治疗AUD的潜在靶点。 改变mRNA甲基化状态。