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

新型 AUTS2 多梳抑制复合物在酒精使用障碍中的作用

基本信息

批准号：
8649699
负责人：
James M Stafford
金额：
$ 3.73万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-30 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8649699
关键词：
AUTS2 gene Accounting Address Affect Affinity Chromatography Alcohol abuse Alcohol consumption Alcohol dependence Alcohols Ataxia Behavior Behavioral Behavioral Genetics Biological Assay Biological Process Biology Brain Brain region ChIP-seq Characteristics Chromatin Complex Consumption DNA Methylation Data Dependence Development Epigenetic Process Ethanol Event Gene Expression Gene Proteins Genes Genetic Genetic Transcription Genetic Variation Histone Acetylation Histones Human Impulsivity Inbred Strain Inbred Strains Mice Intervention Knock-out Knockout Mice Lead Light Link Lysine Maps Mass Spectrum Analysis Measures Modification Molecular Molecular Profiling Mus Neurons PRC1 Protein Pathology Pharmacologic Substance Phenotype Play Polycomb Predisposition Prefrontal Cortex Procedures Prosencephalon RNA Sequences Regulation Regulator Genes Reporter Research Rewards Role Testing Transcriptional Activation Transcriptional Regulation Transgenic Mice Variant Withdrawal Work alcohol use disorder chromatin immunoprecipitation deep sequencing diagnosis design effective therapy fly genome-wide improved in vivo interdisciplinary approach next generation sequencing novel preference prevent problem drinker public health relevance transcriptome sequencing transcriptomics

项目摘要

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.

描述（由申请人提供）：酒精使用障碍（AUD）的一个标志是其异质的遗传和行为基础。下一代测序与转录组学相结合的出现已经开始表明，不仅潜在的基因序列很重要。事实上，关键基因调控的变化（例如表观遗传调控）在 AUD 和相关表型的易感性和发展中发挥着重要作用。这些与 AUD 相关的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性候选基因 2 (AUTS2)。最近，AUTS2 的变异已被证明与多种 AUD 相关行为有关，包括乙醇 (EtOH) 消耗和冲动。初步数据表明 AUTS2 蛋白是转录激活复合物中的关键参与者，这表明 AUTS2 可能在调节 AUD 的基因网络中发挥关键作用。拟议的研究测试了 AUTS2 可能导致与特定 AUD 病理（例如乙醇消耗）相关的异常大脑转录网络的假设。目标 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来鉴定皮质神经元中 AUTS2-染色质复合物的核心成分和染色质动态。目标 2 检查 AUTS2 转录调控在高乙醇偏好和低乙醇偏好小鼠的前额皮质中的作用。在这些研究中，我将检查显示高 (C57BL/6J) 和低 (DBA/2J) EtOH 偏好的近交系小鼠品系前额皮质内 AUTS2 的表达、全基因组占据和转录调控的差异。目标 3 使用转基因小鼠鉴定 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，包括两瓶选择测试，以评估 EtOH 偏好、条件性 EtOH 位置偏好以及 EtOH 诱导的旋转杆共济失调。也许最令人兴奋的可能性是，这些研究将揭示AUTS2可能调节参与AUD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，本研究将指导未来设计针对 AUD 的药物干预措施的研究。