1/1 NADIA U24 Epigenetic/Molecular Scientific Resource Core

摘要 酒精是青春期使用最广泛的成瘾药物之一，持续使用会导致 成年期精神障碍的发展，包括酗酒。表观遗传过程，如 组蛋白乙酰化和DNA甲基化机制，已被证明在神经成熟中发挥作用，通过 有助于大脑发育过程中基因表达的稳定性。青少年的神经生物学 成人饮酒协会(NADIA)发现，青少年间歇性接触乙醇(AIE) 在几个脑回路(前额叶皮质、基底前脑、海马体)产生表观遗传重编程 和杏仁核)，这种情况一直持续到成年，可能是成人精神病理的原因。这些 这些发现促使确定全球表观基因组以确定关键基因内AIE改变的特定基因位点 大脑区域。表观遗传/分子核心的发展是为了评估特定大脑的表观遗传机制 成年期与主要AIE表型相关的区域。表观遗传/分子核心还将开发工具 CRISPR/dCas9方法用于目标基因的表观遗传编辑以直接将AIE诱导的行为联系起来 具有分子机制的表型。核心的目标是提供工具和科学 测试Nadia假设的专业知识，即由于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脑区的转录变化 (前额叶皮质、杏仁核、海马体、下丘脑和基底前脑) 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 建议的工作如下：1)在全基因组水平上检查表观基因组的状态。我们会 进行转座酶可及染色质测序分析(ATAC-SEQ)以确定 Nadia对不同大脑区域的开放和封闭染色质区域进行了研究。核心将提供 AIE期间全球染色质状态的信息到每个组件并识别与 组件的功能研究假设。2)确定AIE诱导的基因特异性表观遗传学 与基因表达变化相关的修饰(组蛋白乙酰化/甲基化/DNA甲基化)。 3)为NAIDA组分提供基因特异性CRISPR/dCas9工具。核心将进行开发和测试 CRISPR/dCas9技术在特定基因位置进行表观遗传编辑(激活或沉默)和 评估功能和行为后果。这些研究将提供更好的理解 跨Nadia的表观遗传重编程对AIE介导的基因表达变化的影响 组件。此外，我们将能够确定可能导致发展的机制 青少年酗酒所致成人精神病理的新治疗策略。

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.