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遗传和行为基础。与转录组学相结合的下一代测序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，表观遗传调节）在AUD和相关表型的易感性和发展中起重要作用。这些基因网络中一个潜在的关键表观遗传调节因子是自闭症易感性候选基因2（AUTS 2）。最近，AUTS 2的变化已被证明与各种AUD相关行为相关，包括乙醇（EtOH）消耗和冲动。结合显示AUTS 2蛋白是转录激活复合物中的关键参与者的初步数据，这表明AUTS 2可能在调节AUD相关基因网络中发挥关键作用。 拟议的研究测试假设AUTS 2可能有助于与选择AUD病理相关的异常脑转录网络（例如，EtOH消耗）。目的1通过纯化、质谱分析以及AUTS 2与组蛋白标记和转录激活的相关性研究，确定AUTS 2-染色质复合物在皮层神经元中的核心组分和染色质动力学。目的2研究AUTS 2转录调控在高和低乙醇偏好小鼠前额叶皮层中的作用。对于这些研究，我将研究在表达，全基因组占用和转录调控的AUTS 2内的近交系小鼠品系，表现出高（C57 BL/6 J）和低（DBA/2 J）乙醇偏好的前额叶皮层的差异。目的3利用转基因小鼠鉴定AUTS 2对AUD的行为和分子贡献。为此，我将产生Auts 2前脑特异性条件性敲除小鼠，并使用行为电池检查EtOH表型，包括两瓶选择测试，以评估EtOH偏好，EtOH的条件性位置偏好，以及EtOH诱导的共济失调。 也许最令人兴奋的可能性是，这些研究将揭示AUTS 2可能调节与AUD相关的大型基因网络，从而阐明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的研究，为AUD设计药物干预措施。

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.