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

16p11.2位点自闭症易感基因对人类干细胞源性神经细胞发育和功能的作用

• 批准号: 10556400
• 负责人: Michael Frederick Wells
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-27 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10556400
• 关键词: 16p11.2; Acceleration; Affect; Attention deficit hyperactivity disorder; Award; Behavior; Biochemical; Biological Models; Brain; Brain Diseases; CRISPR-mediated transcriptional activation; California; Cell Line; Cell model; Cells; Censuses; Child; Cilia; Critical Pathways; DNA; DNA sequencing; Data Set; Defect; Development; Disease; Drug Targeting; Functional disorder; Future; Gene Expression; Genes; Genetic; Goals; High Prevalence; Human; Human Genetics; Impairment; In Vitro; Intellectual functioning disability; Intention; Investigation; Laboratories; Language Disorders; Link; Los Angeles; Maintenance; Malignant Neoplasms; Measures; Mediating; Methods; Modeling; Molecular; Molecular Abnormality; Neurodevelopmental Disorder; Neuroglia; Neuronal Dysfunction; Neurons; Noise; Pathway interactions; Patients; Phase; Phenotype; Population; Prevalence; Process; Proliferating; Protocols documentation; Reporting; Role; SHH gene; Sample Size; Signal Pathway; Susceptibility Gene; Syndrome; System; Techniques; Technology; Testing; Therapeutic Intervention; Tissue-Specific Gene Expression; United States; Universities; Validation; Variant; autism spectrum disorder; brain cell; cell type; cilium biogenesis; diagnostic criteria; effective therapy; empowerment; experimental study; fetal; flasks; genetic risk factor; human fetal brain; human genome sequencing; human model; human stem cells; in vitro Model; in vivo; induced pluripotent stem cell; insight; medical schools; microdeletion; migration; mosaic; nerve stem cell; neurodevelopment; novel; progenitor; repetitive behavior; response; single-cell RNA sequencing; social; stem cell model; stem cells; targeted treatment; transcriptome sequencing

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.

项目概要： 该项目拟议的 R00 阶段将在 Wells 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 名符合自闭症谱系障碍 (ASD)、注意力缺陷多动障碍 (ADHD) 和智力障碍 (ID) 等神经发育障碍的诊断标准。自闭症谱系障碍 (ASD) 的特点是持续的社交障碍、语言缺陷和重复行为，在过去 15 年里增加了 120%，而自闭症谱系障碍 (ASD) 的疾病机制在很大程度上未知且没有针对性的治疗，这一问题进一步加剧。存在治疗干预措施。人类基因组测序的最新进展已开始通过识别几种遗传风险因素来阐明疾病的途径，其中最常见的是 16p11.2 位点 (16p11.2del) 的缺失。初步研究已确定 16p11.2 基因座中神经元功能障碍的特定基因。该提案旨在利用体外诱导多能干细胞 (iPSC) 衍生的人脑细胞来阐明 16p11.2del 表型背后的疾病机制。第一个目标是，我们将尝试在由数十个患者和神经典型对照系组成的神经祖细胞村中使用混合 CRISPR 激活方法来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群患者和对照组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会导致治疗干预的遗传靶点的识别，同时也极大地改变该领域进行人脑疾病体外建模的方式。