1/1 NADIA U24 Epigenetic/Molecular Scientific Resource Core

1/1 NADIA U24 表观遗传学/分子科学资源核心

• 批准号: 10686048
• 负责人: SUBHASH C. PANDEY
• 金额: $ 50.37万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686048
• 关键词: ATAC-seq; Acetylation; Adolescence; Adolescent; Adult; Alcohol abuse; Alcoholism; Alcohols; Amygdaloid structure; Anxiety; Area; Back; Behavioral; Bioinformatics; Biological Assay; Brain; Brain region; CRISPR/Cas technology; Candidate Disease Gene; Chimeric Proteins; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive deficits; DNA Methylation; DNMT3B gene; Data; Development; EP300 gene; Enhancers; Epigenetic Process; Ethanol; Funding; Gene Expression; Genes; Genome; Goals; Hippocampus; Histone Acetylation; Histones; Hypothalamic structure; Individual; Intercistronic Region; Investigation; KDM1A gene; Life; Link; Location; Mediating; Mental Depression; Mental disorders; Messenger RNA; Methylation; Modification; Molecular; Neurobiology; Neurons; Pharmaceutical Preparations; Phenotype; Physiological; Population; Prefrontal Cortex; Psychopathology; Rattus; Research; Research Project Grants; Resources; Risk Factors; Technology; Testing; Time; Transposase; Work; adolescent alcohol exposure; adolescent binge drinking; alcohol use disorder; basal forebrain; behavioral phenotyping; binge drinking; candidate identification; chromatin immunoprecipitation; epigenome; epigenomics; gene network; genomic locus; histone modification; molecular phenotype; novel; novel therapeutic intervention; pharmacologic; promoter; public health relevance; tool; transcriptomics; treatment strategy; underage drinking; whole genome

ABSTRACT Alcohol is one of the most widely used addictive drugs in adolescence, and continued use can lead to the development of psychiatric disorders, including alcoholism, in adulthood. Epigenetic processes, such as histone acetylation and DNA methylation mechanisms, have been shown to play a role in neuromaturation by contributing to the stability of gene expression during brain development. The Neurobiology of Adolescent Drinking in Adulthood (NADIA) consortium has revealed that adolescent intermittent ethanol (AIE) exposure produces epigenetic reprogramming in several brain circuits (prefrontal cortex, basal forebrain, hippocampus and amygdala) that persists until adulthood and might be responsible for adult psychopathology. These findings prompt determination of the global epigenome to determine AIE altered specific gene loci within key brain regions. The Epigenetic/Molecular Core is developed to evaluate epigenetic mechanisms in specific brain regions linked to key AIE phenotypes in adulthood. The Epigenetic/Molecular Core will also develop tools for CRISPR/dCas9 approaches for epigenetic editing of target genes to directly link AIE-induced behavioral phenotypes with molecular mechanisms. The objectives of the Core are to provide tools and scientific expertise to test NADIA’s hypotheses that perturbations of epigenetic targets due to AIE may lead to dynamic changes in epigenetic programming leading to transcriptomic changes in key brain areas (prefrontal cortex, amygdala, hippocampus, hypothalamus, and basal forebrain) which are responsible for persistent behavioral and molecular phenotypes in adulthood. These objectives will be achieved with the following proposed work: 1) To examine the status of the epigenome at the whole genome level. We will perform Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq) to determine the locations of open and closed chromatin domains in different brain regions studied across NADIA. The core will provide information on global chromatin states during AIE to each component and identify “hub” genes related to the component’s hypotheses for functional studies. 2) To determine gene specific AIE-induced epigenetic modifications (histone acetylation/methylation/DNA methylation) associated with changes in gene expression. 3) To provide gene specific CRISPR/dCas9 tools for NADIA components. The core will develop and test CRISPR/dCas9 technologies to make epigenetic editing (activating or silencing) at specific gene locations and evaluate both functional and behavioral consequences. These studies will provide a better understanding of AIE-mediated changes in gene expression via epigenetic reprogramming across NADIA components. In addition, we will be able to identify mechanisms that may lead to the development of novel treatment strategies for adult psychopathologies resulting from adolescent binge drinking.

摘要 酒精是青少年最广泛使用的成瘾药物之一，持续使用会导致 成年后出现精神障碍，包括酗酒。表观遗传过程，如 组蛋白乙酰化和DNA甲基化机制在神经成熟中起作用， 有助于大脑发育过程中基因表达的稳定性。青少年神经生物学 成人饮酒（NADIA）联盟揭示，青少年间歇性乙醇（AIE）暴露 在几个大脑回路（前额叶皮层，基底前脑，海马）中产生表观遗传重编程 和杏仁核），持续到成年，可能是负责成人精神病理学。这些 研究结果提示确定全球表观基因组，以确定AIE改变的关键内的特定基因位点 大脑区域。表观遗传/分子核心是为了评估特定大脑中的表观遗传机制而开发的 与成年期AIE关键表型相关的区域。表观遗传/分子核心还将开发工具， CRISPR/dCas 9方法用于靶基因的表观遗传编辑以直接连接AIE诱导的行为 表型与分子机制。该核心的目标是提供工具和科学 专业知识来测试NADIA的假设，即AIE引起的表观遗传靶点的扰动可能导致 表观遗传编程的动态变化导致关键大脑区域的转录组学变化 （前额叶皮层，杏仁核，海马，下丘脑和基底前脑）， 在成年期持续的行为和分子表型。这些目标将通过以下方式实现： 本文拟开展以下工作：1）在全基因组水平上研究表观基因组的地位。我们将 进行转座酶可重复染色质测序（ATAC-seq）测定，以确定 在NADIA研究的不同大脑区域中开放和闭合的染色质结构域。核心将提供 在AIE过程中，每个组分的全局染色质状态信息，并确定与AIE相关的“枢纽”基因。 功能研究的成分假设。2)为了确定基因特异性AIE诱导的表观遗传 与基因表达变化相关的组蛋白乙酰化/甲基化/DNA甲基化修饰。 3)为NADIA组件提供基因特异性CRISPR/dCas 9工具。核心将开发和测试 CRISPR/dCas 9技术在特定基因位置进行表观遗传编辑（激活或沉默）， 评估功能和行为后果。这些研究将提供一个更好的理解 通过NADIA中的表观遗传重编程，AIE介导的基因表达变化 件.此外，我们将能够确定可能导致发展的机制， 青少年酗酒导致的成人精神病理学的新治疗策略。