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) 是 VA 人群以及一般人群的主要健康问题。 退伍军人管理局使命的一个组成部分是为患有酒精和/或吸毒症状的退伍军人管理局患者提供护理 滥用问题。停止长期摄入乙醇会导致出现负戒断症状 促进饮酒增加和维持酒精成瘾行为的症状。杏仁脑 众所周知，这些结构在焦虑行为和酒精成瘾中发挥着重要作用。这 分子机制，特别是非编码 microRNA（miRNA；21-23 nt）在特定情况下的作用 大脑的神经回路参与乙醇中焦虑样行为的发展 慢性乙醇暴露后的戒断尚不清楚。最近，我们利用微阵列分析 据报道，急性乙醇暴露会在杏仁核中产生差异 miRNA 表达。这个数据 揭示了 miRNA-494 (miR-494) 的表达因急性乙​​醇暴露而降低及其靶标 基因 mRNA 水平增加并参与乙醇抗焦虑作用的调节。这个提议 将扩展这些研究并确定 miR-494 在染色质和突触调节中的作用 杏仁核的重塑，导致酗酒的负面情感状态。此外，该提案将 确定酒精依赖过程中其他 miRNA 表达的变化，然后提出实验建议 研究 miRNA 的这些变化如何协调靶基因通路表达的变化 调节慢性乙醇暴露后戒断期间的焦虑样行为。我们假设 长期接触乙醇后戒断会产生几种 miRNA 的差异表达，并且 其中之一可能是 miR-494。 miR-494 的上调将引起靶基因的变化， 调节杏仁核中的突触可塑性和树突棘，这可能是发育的基础 乙醇戒断期间的焦虑样行为。该提案的具体目标是： 1) 执行 使用微阵列方法对大鼠乙醇戒断期间杏仁核的 miRNA 表达谱进行分析 长期接触乙醇后。新兴的 miRNA 将得到验证，并且其表达会发生变化 假定的目标基因也将被检查。 2) 检查 a) miR-494 antagomir 是否输注在 CeA 中或 急性乙醇挑战会减弱焦虑样行为、表达（mRNA 和蛋白质）缺陷 直接目标（Cbp、p300、Cited2）和间接目标（Bdnf、Arc 和突触素）以及缺陷 慢性乙醇戒断期间大鼠杏仁核突触和树突棘的变化 b) 将 miR-494 的合成模拟物注入对照大鼠的 CeA 中会引起焦虑样症状 行为并减少杏仁核中目标基因的表达。 3) 检查CeA是否有影响 在对照大鼠中输注 Cited2 siRNA 将 a) 产生类似焦虑的行为，从而模仿戒断行为- b) 预防对照大鼠中急性乙醇产生的抗焦虑作用。行为方面 变化将归因于 Cited2 表达的减少以及相关的突触和树突的减少 大鼠大脑脑区的棘。拟议的研究将提供有关 miRNA 调控的新信息 杏仁核中可能参与突触异常的表达及其靶基因 酒精依赖过程中观察到的可塑性，还将确定新的 miRNA 靶标 开发治疗 AUD 的潜在药物。