The role of autism susceptibility genes in the 16p11.2 locus on the development and function of human stem cell-derived neural cells

项目总结： 该项目拟议的R00阶段将在威尔斯实验室进行，该实验室于2021年9月开放，是加州大学洛杉矶分校大卫·格芬医学院人类遗传学系的一部分。最近的报告估计，美国每6名儿童中就有1名符合神经发育障碍的诊断标准，如自闭症谱系障碍(ASD)、注意力缺陷多动障碍(ADHD)和智力残疾(ID)。以持续性社会障碍、语言缺陷和重复行为为特征的自闭症的患病率在过去15年中增加了120%，由于自闭症的发病机制在很大程度上尚不清楚，也没有针对性的治疗干预措施，这一问题进一步加剧。人类基因组测序的最新进展已经开始通过确定几个遗传风险因素来阐明疾病的途径，其中最常见的是16p11.2基因座的缺失(16p11.2del)。初步研究已经提名了16p11.2基因座上与神经元功能障碍有关的特定基因。这项建议旨在利用体外诱导的多能干细胞(IPSC)来源的人脑细胞来阐明16p11.2del表型背后的疾病机制。在第一个目标中，我们将试图使用CRISPR激活方法在由数十个患者和神经典型控制系组成的神经前体细胞村中识别导致疾病相关分子和表型缺陷的16p11.2基因。在第二个目标中，我们将评估和挽救一个由患者和对照组组成的村庄中对主要信号通路激活的异常分子和细胞反应。这些目标的成功完成可能导致确定用于治疗干预的遗传靶点，同时也极大地改变该领域对人类大脑疾病进行体外建模的方式。

Project Summary: The proposed R00 phase of this project will take place in the Wells Laboratory, which opened in September 2021 as part of the Department of Human Genetics in the David Geffen School of Medicine at the University of California Los Angeles. Recent reports estimate that 1 out of every 6 children in the United States meet the diagnostic criteria for neurodevelopmental disorders such as autism spectrum disorders (ASD), attention-deficit hyperactivity disorder (ADHD), and intellectual disability (ID). The prevalence of ASDs, which are characterized by persistent social impairments, language deficits, and repetitive behaviors, has increased by 120% over the past 15 years, a problem further exacerbated by the fact that the disease mechanisms underlying ASDs are largely unknown and no targeted therapeutic interventions exist. Recent progress in human genome sequencing has begun to illuminate pathways to disease through the identification of several genetic risk factors, the most common of which is the deletion of 16p11.2 locus (16p11.2del). Initial studies have nominated specific genes in the 16p11.2 locus in neuronal dysfunction. This proposal aims to elucidate the disease mechanisms underlying 16p11.2del phenotypes using in vitro induced pluripotent stem cell (iPSC)-derived human brain cells. In the first aim, we will attempt to identify the 16p11.2 genes contributing to disease-relevant molecular and phenotypic defects using a pooled CRISPR activation approach in a neural progenitor cell village composed of dozens of patient and neurotypical control lines. In the second aim, we will assess and rescue abnormal molecular and cellular responses to major signaling pathway activation in a village of patients and controls. The successful completion of these aims could lead to the identification of genetic targets for therapeutic intervention, while also dramatically changing the way the field conducts in vitro modeling of human brain disorders.