Cellular signaling in alcoholism

酒精中毒中的细胞信号传导

• 批准号: 10454864
• 负责人: SUBHASH C. PANDEY
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454864
• 关键词: Acetylation; Acute; Addictive Behavior; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcoholism; Alcohols; Amygdaloid structure; Anatomy; Anti-Anxiety Agents; Anxiety; Anxiety Disorders; Attenuated; Behavior; Behavioral; Brain; Brain-Derived Neurotrophic Factor; CREBBP gene; Caring; Cell Nucleus; Chromatin; Chromatin Structure; Chronic; Convulsions; Cytoskeleton; DNA Methylation; Data; Dendritic Spines; Development; Drug Addiction; Drug abuse; E1A-associated p300 protein; EP300 gene; Epigenetic Process; Ethanol; Future; Gene Expression; Gene Expression Regulation; General Population; Genes; Genetic Transcription; Health; Histone Acetylation; Histones; Infusion procedures; Lysine; Medial; Mediating; Messenger RNA; Methylation; MicroRNAs; Microarray Analysis; Mission; Molecular; Motivation; Pathway interactions; Patients; Pharmacotherapy; Play; Population; Post-Transcriptional Regulation; Post-Traumatic Stress Disorders; Proteins; Rattus; Regulation; Reporting; Risk; Role; Signal Transduction; Small Interfering RNA; Structure; Symptoms; Synapses; Synaptic plasticity; Synaptophysin; Testing; Therapeutic Agents; Time; Trans-Activators; Transcription Coactivator; Translational Repression; Tremor; Untranslated RNA; Up-Regulation; Veterans; Withdrawal; Withdrawal Symptom; alcohol effect; alcohol exposure; alcohol use disorder; alcoholism therapy; anxiety-like behavior; base; chromatin remodeling; chronic alcohol ingestion; differential expression; drinking behavior; experimental study; histone modification; miRNA expression profiling; negative affect; neural circuit; novel; prevent; problem drinker; protein complex; public health relevance; recruit

Alcohol use disorder (AUD) is a major health concern in the VA population as well as in the general population. An integral part of the mission of the VA is to provide care for VA patients suffering from alcohol and/or drug abuse problems. Cessation of chronic ethanol consumption leads to development of negative withdrawal symptoms that promote increased alcohol drinking and maintain alcohol addictive behaviors. Amygdaloid brain structures are known to play an important role in anxiety behaviors as well as in alcohol addiction. The molecular mechanisms, and particularly the role of non-coding microRNAs (miRNAs; 21-23 nt), in specific neural circuits of the brain that are involved in the development of anxiety-like behaviors during ethanol withdrawal after chronic ethanol exposure are not well understood. Using microarray analysis, we recently reported that acute ethanol exposure produces differential miRNA expression in the amygdala. This data revealed that expression of miRNA-494 (miR-494) was decreased by acute ethanol exposure and its target gene mRNA levels were increased and involved in the regulation of anxiolytic effects of ethanol. This proposal will extend these studies and establish the role of miR-494 in the regulation of chromatin and synaptic remodeling in the amygdala, leading to negative affective state of alcoholism. In addition, this proposal will identify changes in the expression of other miRNAs during alcohol dependence and then propose experiments to investigate how these changes in miRNAs coordinate shifts in the expression of target gene pathways that regulate anxiety-like behaviors during withdrawal after chronic ethanol exposure. We hypothesize that ethanol withdrawal after chronic exposure will produce differential expression of several miRNAs, and one of these may be miR-494. The upregulation of miR-494 will induce changes in target genes that regulate synaptic plasticity and dendritic spines in the amygdala which may underlie the development of anxiety-like behaviors during ethanol withdrawal. The specific aims of the proposal are: 1) To perform miRNA expression profiling using a microarray approach in the amygdala of rats during ethanol withdrawal after chronic ethanol exposure. The emerging miRNAs will be validated and changes in the expression of their putative target genes will also be examined. 2) To examine if a) miR-494 antagomir infusion in the CeA or acute ethanol challenge will attenuate anxiety-like behaviors, deficits in the expression (mRNA and protein) of direct targets (Cbp, p300, Cited2) and indirect targets (Bdnf, Arc, and synaptophysin) as well as deficits in synapses and dendritic spines in the amygdala of rats during ethanol withdrawal after chronic ethanol exposure and b) synthetic mimics of miR-494 infused into CeA of control rats will provoke anxiety-like behaviors and reduce the expression of target genes in the amygdala. 3) To examine if the effects of CeA infusion of a Cited2 siRNA in control rats will a) produce anxiety-like behaviors, thus mimicking withdrawal- associated anxiety and b) prevent the anxiolytic effects produced by acute ethanol in control rats. Behavioral changes will be due to decreased expression of Cited2 and associated reductions in synapses and dendritic spines in the CeA of rats. The proposed studies will provide new information on the regulation of miRNA expression and their target genes in the amygdala that may be involved in the abnormal synaptic plasticity observed during alcohol dependence, and will also identify novel miRNA targets for the development of potential drugs for the treatment of AUD.

酒精使用障碍（AUD）在VA人群以及普通人群中是一个主要的健康问题。 VA任务不可或缺的一部分是为患有酒精和/或药物的VA患者提供护理 滥用问题。慢性乙醇消耗的停止导致戒断的发展 促进饮酒并维持酒精成瘾行为的症状。杏仁核大脑 众所周知，结构在焦虑行为以及酒精成瘾中起着重要作用。这 分子机制，尤其是非编码microRNA（miRNA; 21-23 NT）的作用 在乙醇期间涉及焦虑症行为发展的大脑的神经回路 慢性乙醇暴露后的戒断尚不清楚。使用微阵列分析，我们最近 报道急性乙醇暴露在杏仁核中产生差异miRNA表达。这个数据 发现miRNA-494（miR-494）的表达通过急性乙醇暴露及其靶标降低 基因mRNA水平增加，并参与乙醇抗焦虑作用的调节。这个建议 将扩展这些研究并确定miR-494在调节染色质和突触中的作用 在杏仁核中进行重塑，导致酗酒的阴性情感状态。此外，该建议将 确定酒精依赖期间其他miRNA表达的变化，然后提出实验 调查miRNA中的这些变化如何在靶基因途径表达的表达中转移 慢性乙醇暴露后戒断期间的焦虑样行为。我们假设这一点 慢性暴露后乙醇撤离将产生几种miRNA的差异表达，并且 其中之一可能是mir-494。 miR-494的上调将引起目标基因的变化 调节杏仁核中的突触可塑性和树突状刺，这可能是发展的基础 乙醇抽出过程中类似焦虑的行为。该提案的具体目的是：1）执行 miRNA表达在乙醇提取过程中使用大鼠杏仁核中的微阵列方法进行分析 慢性乙醇暴露后。新兴的miRNA将得到验证，并在其表达中发生变化 推定的靶基因也将进行检查。 2）检查a）miR-494 CEA中的Antagomir输注还是 急性乙醇挑战将减轻焦虑样行为，表达（mRNA和蛋白质）的缺陷 直接目标（CBP，p300，引用2）和间接目标（BDNF，ARC和突触素）以及缺陷 慢性乙醇后，乙醇戒断期间大鼠杏仁核中的突触和树突状刺 暴露和b）注入控制大鼠CEA的miR-494的合成模仿会引起焦虑样 行为并降低杏仁核中靶基因的表达。 3）检查CEA的影响是否 在对照大鼠中注入引用的2 siRNA将a）产生焦虑样行为，从而模仿戒断 相关的焦虑和b）防止对照大鼠急性乙醇产生的抗焦虑作用。行为 变化将归因于引用2的表达降低以及突触和树突的相关减少 大鼠CEA中的刺。拟议的研究将提供有关miRNA调节的新信息 可能参与异常突触的杏仁核中的表达及其靶基因 在酒精依赖性期间观察到的可塑性，还将确定新颖的miRNA靶标的 开发潜在药物以治疗AUD。