Epigenetic regulation of alcohol tolerance and dependence by methyl CpG binding protein 2

甲基 CpG 结合蛋白 2 对酒精耐受性和依赖性的表观遗传调控

基本信息

批准号：
9133075
负责人：
PIETRO P SANNA
金额：
$ 36.78万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9133075
关键词：
Address Affect Alcohol abuse Alcohol consumption Alcohol dependence Alcohols Automobile Driving Binding Binding Proteins Birds Brain Brain region Chromatin Chronic Complex DNA Dependence Development Down-Regulation Engineering Ethanol Exhibits Experimental Designs Gene Expression Gene Targeting Genes Genetic Transcription Heavy Drinking Impaired cognition Lead Literature Measures Mediating Methyl-CpG-Binding Protein 2 Molecular Molecular Profiling Mus Mutant Strains Mice Pathologic Phenotype Phosphorylation Recording of previous events Recruitment Activity Regulation Regulator Genes Regulon Relapse Rett Syndrome Role Site Substance Addiction Systems Biology Testing Transcription Repressor/Corepressor Treatment Efficacy Validation Viral Vector alcohol effect alcohol exposure alcohol use disorder anxiety-like behavior differential expression drinking epigenetic regulation follow-up gene repression genetic signature genome-wide innovation molecular domain mutant new therapeutic target novel novel therapeutics programs reconstruction sedative social therapeutic target validation studies vapor

项目摘要

Summary Long-term alterations in gene expression programs are believed to be key to the development and progression of alcohol use disorder (AUD). The methyl CpG binding protein 2 (MeCP2), the causative gene of Rett syndrome, is a protein that binds methylated DNA and, in turn, recruits transcriptional repressors resulting in persistent down-regulation of gene expression. We observed that MeCP2 mutant mice with reduced capacity to recruit transcriptional repressors exhibit a robust alcohol–related phenotype characterized by heightened sensitivity to the sedative effects of alcohol, reduced alcohol intake in limited access 2-bottle choice and lack of escalation of drinking after passive induction of dependence. Recent evidence indicates that MeCP2's primary function is to recruit a transcriptional repressor complex at sites of methylated DNA through a discrete molecular domain. Importantly, MeCP2 has been found to regulate a specific set of genes – or regulon – rather than broadly affecting gene expression levels, and we found significant overlap between alcohol-regulated and MeCP2-regulated genes. Thus, in the present project we will test the overarching hypothesis that MeCP2-regulated genes are key to alcohol's effects and to the transition to escalated alcohol drinking in the setting of alcohol dependence. To test the sub-hypothesis that recruitment of transcriptional repressors by MeCP2 is central to its effects on drinking, we will use MeCP2 mutant mice with reduced capacity to recruit transcriptional repressors in comparison with recently introduced MeCP2 mutant mice with increased capacity to recruit transcriptional repressors, to provide optimal perturbation of MeCP2 function for the analysis of MeCP2 regulated gene networks. To test the sub-hypothesis that specific MeCP2 target genes and modulators are key to the transition to escalated drinking associated with alcohol dependence, we will use a state of the art systems biology strategy that we recently validated for the reconstruction and interrogation of genome-wide transcriptional interactomes from brain gene expression profiles. This approach is centered on unbiased identification of transcriptional regulatory relationships from the gene expression effects of the perturbations under study, rather than what is known from the literature or under different sets of perturbations. Rather than identifying long lists of differentially expressed genes, this systems biology strategy identifies and ranks a small number of genes driving the gene signatures associated with the phenotype. Thus, specific mechanistic hypotheses on the role of MeCP2 in the effects of alcohol are obtained, and will then be experimentally validated in paradigms of dependent and non-dependent alcohol drinking. Ultimately, the results of this study will advance our understanding of the molecular mechanisms behind excessive alcohol drinking in the setting of dependence and will lay the rationale for the exploitation of specific MeCP2-regulated genes and modulators for the development of novel therapeutic concepts for AUD.

概括基因表达程序的长期改变被认为是发育和发育的关键。酒精使用障碍 (AUD) 的致病基因甲基 CpG 结合蛋白 2 (MeCP2)。 Rett 综合征是一种结合甲基化 DNA 的蛋白质，进而招募转录抑制因子，从而导致我们观察到 MeCP2 突变小鼠的基因表达持续下调。招募转录抑制因子的能力表现出强大的酒精相关表型，其特征是对酒精镇静作用的批发敏感性，在有限的 2 瓶选择中减少酒精摄入量被动诱导依赖后饮酒没有升级。最近的证据表明 MeCP2 的主要功能是招募转录抑制因子重要的是，我们发现了 MeCP2 通过离散的分子结构域在甲基化 DNA 位点上形成的复合物。调节一组特定的基因（或调节子），而不是广泛影响基因表达水平，我们因此，在本项目中，发现酒精调节基因和 MeCP2 调节基因之间存在显着重叠。我们将检验一个总体假设，即 MeCP2 调节的基因是酒精影响和酒精影响的关键。在酒精依赖的情况下过渡到逐步饮酒以检验子假设： MeCP2 招募转录抑制因子对其对饮酒的影响至关重要，我们将使用 MeCP2 与最近引入的突变小鼠相比，招募转录抑制因子的能力降低 MeCP2 突变小鼠招募转录抑制因子的能力增强，以提供最佳的 MeCP2 功能的扰动用于分析 MeCP2 调节的基因网络以检验子假设。特定的 MeCP2 靶基因和调节剂是向饮酒相关的升级过渡的关键对于酒精依赖，我们将使用我们最近验证的最先进的系统生物学策略从大脑基因表达重建和询问全基因组转录相互作用组该方法的重点是从转录调控关系中公正地识别。正在研究的扰动对基因表达的影响，而不是从文献或文献中已知的该系统不是识别一长串差异表达基因，而是识别不同的扰动组。生物学策略识别并排序少数驱动与相关基因特征的基因因此，获得了关于 MeCP2 在酒精影响中的作用的具体机制假设，然后将在依赖和非依赖饮酒的范例中进行实验验证。最终，这项研究的结果将增进我们对背后分子机制的理解在依赖的情况下过度饮酒，将为利用特定的 MeCP2 调节基因和调节剂用于开发 AUD 的新治疗概念。