Mapping Regulatory Networks of Autism Risk at Cellular Resolution during Neurodevelopment

项目摘要 Muhle博士是一名执业的儿童精神病学家，在发展分子和 用基因工具来阐明疾病的机制。这个职业发展奖的长期目标是 通过整合现状确定自闭症谱系障碍(ASD)潜在的调控网络 ART功能基因组学、单细胞转录组学和精密工程系统生物学分析 早期大脑发育的模型。她在成人和儿童精神病学期间发起了这个项目 耶鲁大学儿童研究中心的研究培训计划，由詹姆斯·努南、博士和 共同指导马修州立大学，医学博士，博士，在努南博士的指导下，世界领先的使用 功能基因组学来了解发育和进化，并得到了道格博士的支持，这是一个无可争议的 作为ASD遗传学的先驱和领导者，Muhle博士发展了调控基因组学和遗传学方面的专业知识 ASD的体系结构。她的职业目标是成为一名独立调查员，研究自闭症患者的角色 染色体结构域解旋酶DNA结合蛋白8(CHD8)在ASD发病中的作用 她的研究目标是确定CHD8的作用以及它的缺失对监管网络的影响。 我们的假设是，CHD8在特定细胞类型中调节ASD风险相关网络，这些细胞类型成为 当CHD8的表达改变时，其表达失调。为了解决这些问题，她成功地生成了一个 CHD8基因敲除小鼠和一只Avi标记的CHD8小鼠，她将用它们来识别CHD8调控的 具有高细胞分辨率的特定细胞类型的网络。她还提出了一种高通量的小分子 筛选影响CHD8调节表达的潜在上游发育信号通路 网络。以下独立的特定目标将解决我们的假设：1)确定细胞类型特定 发育中皮质CHD8单倍体不足引起的表达变化。2)映射CHD8绑定单元格类型 在大脑皮层高分辨率的特定调控网络。3)鉴定CHD8的生物活性调节剂 通过高通量报告实验构建表达网络。我们的理论基础是，通过研究CHD8，一个关键 针对其他ASD风险基因的调控分子，我们将扩大对其他ASD风险基因的了解 并确定CHD8靶向的调控网络，特别是细胞类型，以进一步定义ASD的生物学。 在努南博士的持续指导下，以及Marina Picciotto医学博士的共同指导下， 神经行为障碍的动物模型，与儿童Thomas Fernandez医学博士合作 具有遗传风险因素系统分析专长的精神病学家，以及Smita Krishnaswamy博士， 作为统计学和数学建模前沿领域的计算生物学家，Muhle博士将开发 充分利用她开发的模型系统所需的实验和分析技能。同舟共济 有了耶鲁儿童研究中心的资源和她的职业发展目标，她将实现她的职业生涯 目的阐明脑发育和ASD风险的基本生物学机制。

Project Summary Dr. Muhle is a practicing child psychiatrist with a strong track record of developing molecular and genetic tools to elucidate the mechanism of disease. The long-term goal of this career development award is to identify regulatory networks underlying autism spectrum disorder (ASD) through the integration of state-of-the- art functional genomics, single cell transcriptomics, and systems biology analyses with precision-engineered models of early brain development. She initiated this project during her time in the Adult and Child Psychiatry Research Training Program at the Yale Child Study Center, with the mentorship of James Noonan, Ph.D. and co-mentorship of Matthew State, M.D., Ph.D. With the guidance of Dr. Noonan, a world-wide leader in using functional genomics to understand development and evolution, and the support of Dr. State, an undisputed pioneer and leader in ASD genetics, Dr. Muhle developed expertise in regulatory genomics and the genetic architecture of ASD. Her career goal is to become an independent investigator, examining the role of the ASD risk gene Chromodomain Helicase DNA-binding Protein 8 (CHD8) in the development of ASD. Her research goal is to determine the role of CHD8 and the impact of its loss on regulatory networks. Our hypothesis is that CHD8 regulates ASD risk-associated networks in specific cell-types that become dysregulated when CHD8 expression is altered. To address these questions, she successfully generated a Chd8 constitutive knockout mouse and an Avi-tagged Chd8 mouse that she will use to identify Chd8-regulated networks in specific cell-types at high cellular resolution. She also proposes a high-throughput small molecule screen to explore potential upstream developmental signaling pathways that effect CHD8-regulated expression networks. The following independent specific aims will address our hypothesis: 1) Determine cell-type specific expression changes due to Chd8 haploinsufficiency in the developing cortex. 2) Map Chd8-bound cell-type specific regulatory networks at high resolution in the cortex. 3) Identify bioactive modulators of CHD8 expression networks via high-throughput reporter assay. Our rationale is that by studying CHD8, a key regulatory molecule that targets other ASD risk genes, we will expand our knowledge of other ASD risk genes and identify CHD8-targeted regulatory networks in particular cell-types to further define the biology of ASD. With the continued mentorship of Dr. Noonan and co-mentorship of Marina Picciotto, M.D., an expert in animal models of neurobehavioral disorders, with collaboration of Thomas Fernandez, M.D., a child psychiatrist with expertise in systems analyses of genetic risk factors, and Smita Krishnaswamy, Ph.D., a computational biologist on the cutting-edge of statistical and mathematical modeling, Dr. Muhle will develop the experimental and analytical skills necessary to fully utilize the model systems she has developed. Together with the resources of the Yale Child Study Center and her career development goals, she will attain her career goal to illuminate basic biological mechanisms underlying brain development and risk for ASD.