AUTS2 and the chromatin dynamics of alcohol use disorders

AUTS2 和酒精使用障碍的染色质动力学

基本信息

批准号：
9243734
负责人：
James M Stafford
金额：
$ 16.4万
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：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9243734
关键词：
AUTS2 gene Address Alcohol consumption Alcohols American Animals Architecture Ataxia Behavior Behavioral Biochemical Biochemistry Biological Models Biology Brain Brain region Caliber Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex Consumption DNA Methylation Data Deoxyribonucleases Dependence Development Diagnosis Diagnostic Discipline Disease Disease model Environment Epigenetic Process Ethanol Friends Future Generations Genes Genetic Genetic Transcription Genetic Variation Genomics Goals Histones Human Hypersensitivity Impulsivity Institution Investigation Knock-out Knockout Mice Learning Life Link Literature Lysine Measures Mental disorders Mentorship Methods Modeling Molecular Molecular Biology Molecular Biology Techniques Molecular Conformation Molecular Profiling Mus Neurobiology Neurons Pathology Pharmacologic Substance Pharmacology Phase Phenotype Plant Roots Play Prefrontal Cortex Prevention Procedures Prosencephalon Research Research Personnel Resources Rewards Rodent Role Self Administration Shapes Substance abuse problem System Techniques Testing Training Transcriptional Activation Transcriptional Regulation Variant Withdrawal Work alcohol abuse therapy alcohol intervention alcohol research alcohol use disorder career chromatin immunoprecipitation chromatin remodeling collaborative environment deep sequencing diagnosis design effective therapy epigenetic regulation experience genome editing genome-wide histone modification improved in vivo innovation interdisciplinary approach member mouse model novel post-doctoral training preference prevent problem drinker programs relating to nervous system research and development tool transcriptome sequencing

项目摘要

Project Summary Candidate My overarching goal is to bridge the gap between basic discoveries and the prevention and treatment of alcohol use disorders (AUD). Towards accomplishing that goal I have sought career and scientific opportunities that expose me to a variety of approaches to tackling AUD from molecular, behavioral and neural system perspectives. The major emphasis of my graduate training was developing sophisticated behavioral procedures that facilitate pharmacological manipulations and investigation of the molecular underpinnings of psychiatric and AUD. To unite this training with the deep biochemistry and molecular tools required to determine how epigenetic mechanisms contribute to AUD, I sought my post-doctoral training and research in the lab of Dr. Reinberg, who is a world-renowned leader in chromatin biology and transcription. My ultimate goal for the R00 phase is to become an independent investigator and tackle the root, epigenetic causes of AUD and make a large impact on their diagnosis, prevention and treatment. I plan to accomplish this by bridging disciplines so that I can approach AUD from molecular, behavioral and neural system perspectives. This K99/R00 application represents only the beginning of this work and I am most excited by its potential to discover novel molecular mechanisms in AUD through my genome-wide and biochemical approaches. Environment Dr. Reinberg's lab is a fantastic place to gain the experience I need to launch my future career as an independent investigator. Under his mentorship and through his dynamic lab, I will learn a host of sophisticated biochemistry and molecular biology techniques. Importantly, this lab is well established and has deep resources for probing challenging questions in chromatin biology. These well-established resources and expertise include all of the biochemistry and molecular biology techniques required for training in the studies I propose. One of the truly remarkable things about the Reinberg lab is the high caliber and deep expertise of its members working on a number of challenging questions requiring the development of innovative techniques and approaches. The environment in his lab dovetails nicely with the supportive environment at my host institution, my collaborators and the training that is sure to enrich my experience and propel my professional development Research Alcohol (ethanol, EtOH) use disorders (AUD) are characterized by heterogeneous genetic and behavioral underpinnings. Indeed, growing evidence suggests that disruptions in the underlying gene sequence can only partially account for the molecular profile of AUD. An emerging theme is that the expression profile of large gene networks is markedly altered in the alcoholic brain. Preliminary data and recent literature suggest that autism susceptibility candidate 2 (AUTS2) may represent one such key epigenetic regulator that drives aberrant transcriptional programs in AUD. This proposal examines how AUTS2 and associated chromatin dynamics drive transcriptional changes that may underlie specific AUD phenotypes. AIM1 will first identify the core components and chromatin dynamics of AUTS2-related complexes directly in C57Bl/6J (C57) neurons using a sophisticated biochemistry and molecular biology approach. Secondly, AIM1 will probe how these AUTS2-associated chromatin dynamics impact transcriptional regulation in the PFC of mice selectively bred for high and low EtOH preference (HAP and LAP mice, respectively) through ChIP-seq and RNA-seq. AIM 2 examines how the expression of AUTS2 itself is regulated in both mouse AUD models (i.e., HAP vs. LAP) and human neural culture that models an AUTS2 SNP associated with EtOH consumption and AUTS2 expression. AIM 3 identifies behavioral and molecular contributions of AUTS2 to AUD-associated behaviors in vivo through the generation of an inducible, Auts2-conditional knockout (CKO) mouse. Through this work, I hope to reveal how AUTS2 drives AUD transcriptional and behavioral phenotypes, providing guides for diagnostic and targeted interventions of AUD.

项目概要候选人我的首要目标是弥合基本发现与预防和治疗之间的差距酒精使用障碍（AUD）。为了实现这个目标，我寻求职业和科学这些机会让我接触到从分子、行为和神经方面解决 AUD 问题的各种方法系统视角。我的研究生培训的主要重点是培养复杂的行为促进药理学操作和分子基础研究的程序精神科和澳元。将这种培训与深入的生物化学和分子工具结合起来确定表观遗传机制如何促进 AUD，我寻求博士后培训和研究 Reinberg 博士的实验室是世界著名的染色质生物学和转录领域的领军人物。我的终极 R00 阶段的目标是成为一名独立研究者并解决问题的根本原因、表观遗传原因 AUD 对其诊断、预防和治疗产生很大影响。我计划通过以下方式完成此任务跨学科的桥梁，使我能够从分子、行为和神经系统的角度来研究 AUD。这个 K99/R00 应用程序仅代表这项工作的开始，我对它的潜力感到非常兴奋通过我的全基因组和生化方法发现 AUD 的新分子机制。环境 Reinberg 博士的实验室是一个绝佳的地方，可以让我获得开启未来职业生涯所需的经验。独立调查员。在他的指导下，通过他充满活力的实验室，我将学到许多复杂的知识生物化学和分子生物学技术。重要的是，该实验室设施完善，具有深厚的用于探索染色质生物学中具有挑战性的问题的资源。这些完善的资源和专业知识包括研究培训所需的所有生物化学和分子生物学技术提出。 Reinberg 实验室真正引人注目的事情之一是其高素质和深厚的专业知识成员致力于解决一些需要开发创新技术的挑战性问题和方法。他实验室的环境与我东道主的支持环境非常吻合机构、我的合作者和培训必将丰富我的经验并推动我的专业发展发展研究酒精（乙醇，EtOH）使用障碍（AUD）的特点是异质遗传和行为基础。事实上，越来越多的证据表明，潜在基因序列的破坏只能部分解释了 AUD 的分子特征。一个新兴的主题是大的表达轮廓酗酒者大脑中的基因网络发生了显着改变。初步数据和最近的文献表明自闭症易感性候选 2 (AUTS2) 可能代表一种关键的表观遗传调节因子，它驱动 AUD 中的异常转录程序。该提案研究了 AUTS2 和相关染色质如何动力学驱动的转录变化可能是特定 AUD 表型的基础。 AIM1首先会识别 C57Bl/6J (C57) 神经元中 AUTS2 相关复合物的核心成分和染色质动力学使用复杂的生物化学和分子生物学方法。其次，AIM1 将探讨这些 AUTS2 相关染色质动态影响选择性培育小鼠 PFC 的转录调控通过 ChIP-seq 和 RNA-seq 进行高和低 EtOH 偏好（分别为 HAP 和 LAP 小鼠）。目标2 检查 AUTS2 本身的表达在两种小鼠 AUD 模型（即 HAP 与 LAP）中是如何受到调节的人类神经培养物，模拟与 EtOH 消耗和 AUTS2 表达相关的 AUTS2 SNP。 AIM 3 通过以下方式识别 AUTS2 对体内 AUD 相关行为的行为和分子贡献诱导型 Auts2 条件敲除 (CKO) 小鼠的产生。通过这个作品，我希望揭示 AUTS2 如何驱动 AUD 转录和行为表型，为诊断和治疗提供指南澳元有针对性的干预措施。