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）的一个标志是其异质性的遗传和行为基础。下一代测序结合转录组学的出现已经开始揭示，不仅仅是潜在的基因序列是重要的。事实上，关键基因调控的变化（如表观遗传调控）在AUD的易感性和发展及相关表型中起着重要作用。自闭症易感性候选基因2 （AUTS2）是这些基因网络中一个潜在的关键表观遗传调控因子。最近，AUTS2的变异已被证明与多种与aud相关的行为有关，包括乙醇（EtOH）消耗和冲动。结合初步数据显示AUTS2蛋白在转录激活复合体中起关键作用，这表明AUTS2可能在调节AUD的基因网络中起关键作用。拟议的研究验证了AUTS2可能导致与特定AUD病理（例如，EtOH消耗）相关的异常脑转录网络的假设。目的1将通过纯化、质谱分析以及AUTS2与组蛋白标记和转录激活的关联，鉴定皮质神经元中AUTS2-染色质复合物的核心成分和染色质动力学。目的2研究了AUTS2转录调控在高etoh偏好和低etoh偏好小鼠前额叶皮层中的作用。在这些研究中，我将研究在表现出高（C57BL/6J）和低（DBA/2J） EtOH偏好的近交小鼠品系中，AUTS2在前额叶皮层内的表达、全基因组占用和转录调控的差异。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前脑特异性条件敲除小鼠，并使用行为电池检查EtOH表型，包括评估EtOH偏好的两瓶选择测试，EtOH的条件位置偏好，以及EtOH诱导的旋转杆上的失调。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相关的大型基因网络，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。通过研究这些分子事件如何映射到AUD，本研究将指导未来设计AUD药物干预措施的研究。

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.