The role of a novel AUTS2 polycomb repressive complex in alcohol use disorders

描述(由申请人提供)：酒精使用障碍(AUD)的一个特征是其不同的遗传和行为基础。下一代测序与转录组学相结合的出现已经开始揭示，不仅潜在的基因序列是重要的。事实上，关键基因调控的变化(例如，表观遗传调控)在AUD及其相关表型的易感和发展中发挥着重要作用。参与AUD的这些基因网络的一个潜在的关键表观遗传调节因子是自闭症易感候选2基因(AUTS2)。最近，AUTS2的变异被证明与一系列与AUD相关的行为有关，包括酒精(Etoh)消费和冲动。结合初步数据显示，AUTS2蛋白是转录激活复合体中的关键角色，这表明AUTS2可能在调节AUD的基因网络中发挥关键作用。这项拟议的研究测试了假设AUTS2可能有助于与特定的AUD病理相关的异常大脑转录网络(例如，乙醇消耗)。目的1通过纯化、质谱法以及AUTS2与组蛋白标记和转录激活的关系，鉴定皮质神经元AUTS2-染色质复合体的核心成分和染色质动力学。目的2研究AUTS2转录调控在高乙醇偏好和低乙醇偏好小鼠前额叶皮质中的作用。对于这些研究，我将检验AUTS2在近交系小鼠前额叶皮质中的表达、全基因组占有率和转录调控的差异，这些品系表现出高(C57BL/6J)和低(DBA/2J)乙醇偏好。目的3利用转基因小鼠鉴定AUTS2对AUD的行为和分子贡献。为此，我将生成一只AUTS2前脑特异的条件性基因敲除小鼠，并使用行为电池检测乙醇表型，包括两瓶选择测试以评估乙醇偏好、对乙醇的条件性位置偏好以及乙醇诱导的旋转棒共济失调。也许最令人兴奋的可能性是，这些研究将揭示AUTS2可能调节与AUD有关的大型基因网络，从而揭示基因网络的大规模破坏如何导致AUD表型的出现和易感性。通过研究这些分子事件如何映射到AUD，这项研究将指导未来设计AUD药物干预的研究。

DESCRIPTION (provided by applicant): A hallmark of alcohol use disorders (AUD) is its heterogeneous genetic and behavioral underpinnings. The advent of next generation sequencing combined with transcriptomics has begun to reveal that not only the underlying gene sequence is important. Indeed, changes in the regulation of key genes (e.g., epigenetic regulation) play a strong role in the predisposition to and development of AUD and associated phenotypes. One potential key epigenetic regulator of these gene networks involved in AUD is the autism susceptibility candidate 2 gene (AUTS2). Recently, variations of AUTS2 have been shown to be associated with a variety of AUD-related behaviors including ethanol (EtOH) consumption and impulsivity. Combined with preliminary data showing that the AUTS2 protein is a critical player in a transcriptional activation complex, this suggests that AUTS2 may play a key role in regulating gene networks underlying AUD. The proposed research tests the hypothesis AUTS2 may contribute to aberrant brain transcriptional networks associated with select AUD pathologies (e.g., EtOH consumption). Aim 1 will identify the core components and chromatin dynamics of AUTS2-chromatin complex in cortical neurons through purification, mass spectrometry as well as AUTS2 association with histone marks and transcriptional activation. Aim 2 examines the role of AUTS2 transcription regulation in the prefrontal cortex of high- and low-EtOH preferring mice. For these studies I will examine differences in the expression, genome-wide occupancy and transcriptional regulation of AUTS2 within the prefrontal cortex of inbred mouse strains that show high (C57BL/6J) and low (DBA/2J) EtOH preference. Aim 3 identifies behavioral and molecular contributions of AUTS2 to AUD using transgenic mice. For this I will generate an Auts2 forebrain-specific conditional knock-out mouse and examine EtOH phenotypes using a behavioral battery including a two-bottle choice test to assess EtOH preference, conditioned place preference for EtOH, and EtOH-induced ataxia on the rotarod. Perhaps the most exciting possibility is that these studies will reveal that AUTS2 may regulate large gene networks involved in AUD thus shedding light on how large-scale disruptions in gene networks lead to the emergence of and predisposition to AUD-phenotypes. By examining how these molecular events map onto AUD, this study will guide future research in devising pharmaceutical interventions for AUD.