Mapping Regulatory Networks of Autism Risk at Cellular Resolution during Neurodevelopment

项目摘要 穆勒博士是一位执业儿童精神病学家，在发展分子和 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 通过整合自闭症谱系障碍（ASD）的现状， 功能基因组学、单细胞转录组学和系统生物学分析， 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 耶鲁儿童研究中心的研究培训项目，由James Noonan博士指导。和 马修州立大学医学博士的共同导师博士在努南博士的指导下，努南博士是使用 功能基因组学，以了解发展和进化，以及博士的支持状态，一个无可争议的 作为ASD遗传学的先驱和领导者，穆勒博士在调控基因组学和遗传学方面积累了丰富的经验。 ASD的架构。她的职业目标是成为一名独立调查员，研究ASD的作用 染色体结构域解旋酶DNA结合蛋白8（CHD8）在ASD发展中的作用 她的研究目标是确定CHD8的作用以及其损失对监管网络的影响。 我们的假设是，CHD8在特定的细胞类型中调节ASD风险相关网络， 当CHD8的表达改变时，为了解决这些问题，她成功地生成了一个 Chd8组成型敲除小鼠和Avi标记的Chd8小鼠，她将使用它们来鉴定Chd8调节的 以高细胞分辨率在特定细胞类型中的网络。她还提出了一种高通量的小分子 筛选以探索影响CHD8调节表达的潜在上游发育信号通路 网络.以下独立的具体目标将解决我们的假设：1）确定细胞类型特异性 由于发育中的皮质中Chd8单倍不足而引起的表达变化。2)映射Chd8结合细胞类型 大脑皮层中的高分辨率的特定调控网络。3)鉴定CHD8的生物活性调节剂 通过高通量报告基因测定的表达网络。我们的理由是，通过研究CHD8， 针对其他ASD风险基因的调节分子，我们将扩大我们对其他ASD风险基因的了解 并鉴定特定细胞类型中CHD8靶向的调控网络，以进一步确定ASD的生物学。 在努南博士的持续指导和玛丽娜·皮乔托医学博士的共同指导下，的专家 神经行为障碍的动物模型，与医学博士托马斯费尔南德斯合作，一个孩子 具有遗传风险因素系统分析专长的精神病学家，和Smita Krishnaswamy博士，一 作为统计和数学建模前沿的计算生物学家，穆勒博士将开发 充分利用她开发的模型系统所需的实验和分析技能。一起 凭借耶鲁儿童研究中心的资源和她的职业发展目标，她将实现自己的职业生涯 目的是阐明大脑发育和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.