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

• 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
• 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

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.

摘要 基因表达程序的长期改变被认为是发展和 酒精使用障碍的进展(AUD)。甲基CpG结合蛋白2(MeCP2)，致病基因 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导致 持续下调基因表达。我们观察到MeCP2突变小鼠具有减少的 招募转录抑制因子的能力表现出与酒精相关的强健表型，其特征是 提高了对酒精镇静作用的敏感性，减少了酒精摄入量，提供了两瓶限量选择 在被动地诱导依赖后，饮酒没有升级。 最近的证据表明，MeCP2的S的主要功能是招募一个转录抑制因子 通过一个离散的分子结构域在甲基化DNA的位置上的复合体。重要的是，已经发现了MeCP2 来调控一组特定的基因或调节子，而不是广泛地影响基因表达水平，我们 发现酒精调控的基因和MeCP2调控的基因有显著重叠。因此，在本项目中， 我们将测试最重要的假设，即MeCP2调节的基因是酒精影响的关键，以及 在酒精依赖的背景下向饮酒升级过渡。来验证子假说 MeCP2对转录抑制子的招募是其对饮酒影响的核心，我们将使用MeCP2 与最近引进的小鼠相比，突变小鼠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 MeCP2调控基因网络分析中MeCP2函数的扰动。为了验证子假设 特定的MeCP2靶基因和调节器是过渡到与酗酒相关的升级的关键 对于酒精依赖，我们将使用一种最先进的系统生物学策略，这是我们最近验证的 从脑基因表达重建和查询全基因组转录相互作用 配置文件。这种方法的核心是公正地识别转录调控关系 研究中的扰动对基因表达的影响，而不是从文献中或以下已知的 不同的扰动集合。这一系统不是识别一长串差异表达的基因，而是 生物策略识别并排序了少数驱动与 表型。因此，获得了关于MeCP2在酒精效应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，这项研究的结果将促进我们对背后的分子机制的理解。 在过度饮酒的背景下依赖并将为具体的剥削奠定理论基础 MeCP2调控的基因和调节剂，用于开发AUD的新治疗概念。