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人群和普通人群的主要健康问题。 VA的使命的一个组成部分是为患有酒精和/或药物的VA患者提供护理 虐待问题停止慢性乙醇消耗导致负戒断的发展 促进饮酒增加并维持酒精成瘾行为的症状。杏仁样脑 已知结构在焦虑行为以及酒精成瘾中起重要作用。的 分子机制，特别是非编码microRNAs（miRNAs; 21-23 nt），在特异性 大脑的神经回路，参与乙醇过程中焦虑样行为的发展 慢性乙醇暴露后的戒断症状尚不清楚。利用微阵列分析，我们最近 报道，急性乙醇暴露在杏仁核中产生差异的miRNA表达。该数据 结果显示，急性乙醇暴露降低了miRNA-494（miR-494）的表达，其靶点是： 基因mRNA水平增加，并参与调节乙醇的抗焦虑作用。这项建议 将扩展这些研究，并建立miR-494在染色质和突触调节中的作用。 杏仁核的重塑，导致酒精中毒的负面情绪状态。此外，该提案将 确定酒精依赖期间其他miRNAs表达的变化，然后提出实验 为了研究这些miRNAs的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 慢性暴露后的乙醇戒断会产生几种miRNA的差异表达， 其中之一可以是miR-494。miR-494的上调将诱导靶基因的变化， 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。 在酒精戒断过程中的焦虑行为。该提案的具体目标是：1）执行 乙醇戒断大鼠杏仁核miRNA表达谱的微阵列研究 慢性酒精暴露后。新出现的miRNAs将被验证，其表达的变化将被证实。 还将检查推定的靶基因。2)检查a）在CeA中输注miR-494厄洛莫或 急性乙醇攻击将减弱焦虑样行为， 直接靶点（Cbp，p300，Cited 2）和间接靶点（Bdnf，Arc和突触素）以及 慢性乙醇戒断大鼠杏仁核内突触和树突棘的变化 暴露和B）注入对照大鼠CeA的miR-494的合成模拟物将引起焦虑样 行为，并减少杏仁核中靶基因的表达。3)为了检验CeA的作用 在对照大鼠中输注Cited 2 siRNA将a）产生焦虑样行为，从而模拟戒断- 相关的焦虑和B）防止对照大鼠中急性乙醇产生的抗焦虑作用。行为 这些变化可能是由于Cited 2表达减少以及突触和树突状细胞中相关的减少。 大鼠脑前区的棘。这些研究将为miRNA的调控提供新的信息。 杏仁核中的表达及其靶基因，可能参与异常突触 在酒精依赖过程中观察到的可塑性，也将确定新的miRNA靶点， 开发治疗AUD的潜在药物。