AUTS2 and the chromatin dynamics of alcohol use disorders

项目摘要 侯选人 我的首要目标是弥合基本发现和预防和治疗之间的差距 酒精使用障碍(AUD)。为了实现这一目标，我一直在寻求职业和科学 让我接触到从分子、行为和神经方面处理AUD的各种方法的机会 系统视角。我研究生训练的主要重点是培养成熟的行为 促进药物操作和分子基础研究的程序 精神疾病和澳元。将这种培训与深度生物化学和分子工具相结合 确定表观遗传机制如何有助于AUD，我寻求博士后培训和研究 莱因伯格博士的实验室，他是染色质生物学和转录领域的世界知名领导者。我的终极目标 R00阶段的目标是成为一名独立的调查者，并解决 并对其诊断、预防和治疗产生较大影响。我计划通过以下方式实现这一目标 在学科之间架起桥梁，这样我就可以从分子、行为和神经系统的角度来研究澳大利亚大学。 这个K99/R00应用程序只是这项工作的开始，我最兴奋的是它的潜力 通过我的全基因组和生物化学方法，发现AUD的新分子机制。 环境 莱因伯格博士的实验室是一个极好的地方，可以让我获得启动我未来职业生涯所需的经验 独立调查员。在他的指导下，通过他的动态实验室，我将学习到许多复杂的 生物化学和分子生物学技术。重要的是，这个实验室建立得很好，而且有很深的 探讨染色质生物学中具有挑战性的问题的资源。这些成熟的资源和 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 求婚。莱因伯格实验室真正了不起的事情之一是其高素质和深厚的专业知识 成员致力于一些具有挑战性的问题，需要开发创新的技术 和接近。他实验室的环境与我主人的支持性环境非常吻合。 机构，我的合作者，以及一定会丰富我的经验和推动我的专业的培训 发展 研究 酒精(乙醇)使用障碍(AUD)的特点是遗传和行为的异质性 支撑点。事实上，越来越多的证据表明，潜在基因序列的中断只会 部分解释了AUD的分子轮廓。一个新兴的主题是，大型 在酗酒的大脑中，基因网络发生了显著的变化。初步数据和最近的文献表明 自闭症易感性候选基因2(AUTS2)可能代表了这样一个关键的表观遗传调控因子，它推动 AUD的异常转录程序。这项建议研究了AUTS2和相关的染色质 动力学驱动转录变化，这些变化可能是特定AUD表型的基础。AIM1将首先识别 直接在C57BL/6J(C57)神经元中AUTS2相关复合体的核心成分和染色质动力学 使用先进的生物化学和分子生物学方法。其次，AIM1将调查这些 AUTS2相关染色质动力学影响选择性培养小鼠PFC的转录调控 高和低乙醇偏好(分别为HAP和LAP小鼠)通过ChIP-seq和RNA-seq。目标2 研究了AUTS2本身在两种小鼠AUD模型(即，HAP与LAP)中的表达是如何调节的 人类神经培养，模拟与乙醇消耗和AUTS2表达相关的AUTS2 SNP。 目的3确定AUTS2在体内对AUD相关行为的行为和分子贡献 可诱导的AUTS2条件性基因敲除(CKO)小鼠的产生。通过这部作品，我希望揭示 AUTS2如何驱动AUD转录和行为表型，为诊断和 澳元的针对性干预。

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.