Connecting gene regulatory mechanisms in human brain to psychiatric illness

项目摘要/摘要 越来越多的证据表明，染色质调节器的异常与一系列神经精神疾病有关， 包括自闭症和精神分裂症。然而，这些染色质调节剂如何影响心脏的病理生理学 精神疾病目前还没有得到很好的理解。这项提议试图描述全基因组的特征 人类大脑中代表出生前和出生后两个主要时期的染色质景观 通过Hi-C，一种全基因组的染色体构象捕获技术，进行发育阶段的研究。蜂窝 异质性将通过荧光激活的核分类(FANS)来描绘神经元和非神经元 神经细胞分开。总而言之，我们将确定发育和细胞类型特定的三维 (3D)作为基因调控基础的染色质结构(目标1)。我们将利用染色质相互作用 识别与易感基因的非编码变异体发生物理作用的基因 神经精神障碍(目标2)。发育和细胞类型特异性3D染色质的研究 建筑学将规定发育阶段和细胞类型，这对人体的病理生理学有贡献。 精神错乱。此外，交叉紊乱分析将描绘共同的遗传病因学和神经生物学 整个精神疾病的机制。非编码变体与Hi-C的函数关系 确定的目标基因将通过使用大规模平行报告分析的实验验证来询问 (MPRA)和CRISPR/Cas9介导的基因组编辑(目标3)。总体而言，该项目的完成将 解开精神病学病因学背后的基因组折叠原理。 这一奖项的K99阶段将是我完成博士后培训的关键。它将允许我 继续在丹尼尔·H·格施温德博士的监督下工作，最终确定目标1和目标2，他的知识 神经发育生物学和遗传学的研究将是有益的。最重要的是，和其他三个人一起训练 我想以独立的身份建立的实验需要我的顾问委员会的成员 研究员。Jason Ernst博士将帮助我完成目标1和目标2中的综合分析，以及 MPRA在AIM 3中的结果。Eleazar Ekin博士将指导我在关联研究中识别因果变异 目标2中的人类精神障碍。斯里·科苏里博士将在目标3中帮助我设计和分析MPRA。 最后，加州大学洛杉矶分校的环境和机会(研讨会、会议和会议)将提供 为我博士后培训的最后阶段做好准备。

Project Summary/Abstract Growing evidence implicates abnormalities in chromatin regulators in a range of neuropsychiatric illnesses, including ASD and schizophrenia. However, how these chromatin regulators affect the pathophysiology of psychiatric illnesses is currently not well understood. This proposal seeks to characterize genome-wide chromatin landscape in human brains across two major epochs representing prenatal and postnatal developmental stages via Hi-C, a genome-wide chromosome conformation capture technology. Cellular heterogeneity will be delineated via fluorescence-activated nuclear sorting (FANS) to profile neurons and non- neural cells separately. Collectively, we will determine the developmental and cell-type specific 3-dimensional (3D) chromatin architecture that underlies gene regulation (Aim 1). We will leverage chromatin interaction maps to identify genes that are physically interacting with non-coding variants that predispose to neuropsychiatric disorders (Aim 2). Investigation of developmental and cell-type specific 3D chromatin architecture will specify the developmental stage and cell type that contributes to the pathophysiology of the disorders. Moreover, cross-disorder analysis will delineate shared genetic etiology and neurobiological mechanisms across psychiatric illnesses. The functional relationship between non-coding variants and Hi-C defined target genes will be interrogated by experimental validation using massive parallel reporter assays (MPRA) and CRISPR/Cas9-mediated genome editing (Aim 3). Collectively, completion of this project will unravel the folding principles of the genome underlying the psychiatric etiology. The K99 phase of this award will be essential for me to finalize my postdoctoral training. It will allow me to continue to work under supervision of Dr. Daniel H. Geschwind to finalize Aims 1 and 2, where his knowledge of neurodevelopmental biology and genetics will be instrumental. Most importantly, training with the other three members of my advisory board will be required for the experiments I want to set up as an independent researcher. Dr. Jason Ernst will help me with the integrative analysis in Aims 1 and 2, and the analysis of MPRA results in Aim 3. Dr. Eleazar Eskin will guide me in identifying causal variants in association studies of human psychiatric disorders in Aim 2. Dr. Sri Kosuri will aid me with the design and analysis of MPRA in Aim 3. Finally, the environment and opportunities at UCLA (workshops, conferences, and meetings) will provide the ideal setting for the final stages of my postdoctoral training.