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: 9922991
• 负责人: Michael Frederick Wells
• 金额: $ 10.23万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922991
• 关键词: 16p11.2; Affect; Animal Model; Attention deficit hyperactivity disorder; Award; Behavior; Biochemical; Bioinformatics; Biological Models; Brain; Brain Diseases; Cell Line; Cell model; Cells; Censuses; Child; Cilia; Communication; Critical Pathways; DNA; DNA sequencing; Data; Data Set; Defect; Development; Development Plans; Diagnostic; Disease; Disease model; Drug Targeting; Experimental Designs; Feedback; Foundations; Functional disorder; Future; Gene Expression; Genes; Genetic; Goals; Grant; Growth; High Prevalence; Human; Impairment; In Vitro; Institutes; Institution; Intellectual functioning disability; Intention; International; Investigation; Laboratories; Laboratory Research; Language Disorders; Lead; Link; Maintenance; Malignant Neoplasms; Measures; Mediating; Mentors; Methods; Modeling; Molecular; Mosaicism; Mus; Neurodevelopmental Disorder; Neuroglia; Neuronal Dysfunction; Neurons; Noise; Pathway interactions; Patients; Phase; Phenotype; Population; Prevalence; Principal Investigator; Process; Protocols documentation; Reporting; Research; Role; SHH gene; Sample Size; Signal Pathway; Susceptibility Gene; Syndrome; System; Techniques; Technology; Testing; Therapeutic Intervention; Tissue-Specific Gene Expression; Training; Type I Epithelial Receptor Cell; United States; Universities; Validation; Variant; Work; Writing; Zebrafish; autism spectrum disorder; base; brain cell; career; career development; cell type; cilium biogenesis; effective therapy; experimental study; fetal; flasks; genetic risk factor; human fetal brain; human fetal brain tissue; human genome sequencing; human model; human stem cells; improved; in vitro Model; in vivo; induced pluripotent stem cell; innovation; insight; meetings; microdeletion; migration; nerve stem cell; neurodevelopment; novel; progenitor; repetitive behavior; single-cell RNA sequencing; skills; social; stem cells; symposium; targeted treatment; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT 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, though these findings are built on mouse and zebrafish models rather than human neural cell types. 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 Aim 1 (K99), human iPSC- derived neural progenitor cells and neurons generated using novel protocols will be compared to human fetal brain tissue using single-cell RNA sequencing (scRNA-seq) techniques to validate these in vitro cellular models for future studies. Aim 2 of this proposal (K99) will employ an innovative “population-in-a-dish” strategy in which stem cell lines from many different neurotypical and 16p11.2del patients will be pooled into one culture flask to interrogate phenotypic differences. Aim 3 (R00) will leverage these scRNA-seq techniques to decipher the role of 16p11.2 genes in specific pathways important for neurodevelopment. 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. These experiments will provide new training for the principal investigator (PI) of this proposal in scRNA-seq and bioinformatics methods that will serve as the foundation of an independent research laboratory that will use stem cell-derived neural cells and large transcriptome datasets, combined with animal models, to elucidate the cellular and molecular mechanisms governing neurodevelopmental disorders. This work will be completed at the Broad Institute and Harvard University, where the opportunities for technical and intellectual growth are innumerable. The PI will attend regular meetings with mentors and collaborators to receive feedback on experimental design and career decisions. The PI will attend grant writing and project management courses at Harvard, while also improving his communication skills by presenting data at international scientific conferences. As a whole, this career development plan will help the PI establish his own group at a research-oriented academic institution and become a leader in the field of neurodevelopmental disease modeling.

项目概要/摘要 最近的报告估计，美国每 6 名儿童中就有 1 名符合以下诊断标准： 神经发育障碍，例如自闭症谱系障碍 (ASD)、注意力缺陷多动障碍 (ADHD) 和智力障碍 (ID) 的患病率，其特点是持续的社交。 障碍、语言缺陷和重复行为在过去 15 年中增加了 120%， 自闭症谱系障碍 (ASD) 的疾病机制在很大程度上尚不清楚，并且这一事实进一步加剧了这一问题 目前，人类基因组测序已开始取得进展。 通过识别几种遗传风险因素（最常见的）来阐明疾病的途径 这是 16p11.2 基因座的缺失（16p11.2del）。初步研究已经提名了 16p11.2 中的特定基因。 神经元功能障碍的位点，尽管这些发现是基于小鼠和斑马鱼模型而不是人类 该提案旨在阐明 16p11.2del 表型的疾病机制。 使用体外诱导多能干细胞 (iPSC) 衍生的人脑细胞 In Aim 1 (K99)，人 iPSC-。 使用新方案生成的衍生神经祖细胞和神经元将与人类胎儿进行比较 使用单细胞 RNA 测序 (scRNA-seq) 技术对脑组织进行验证，以验证这些体外细胞模型 该提案的目标 2（K99）将采用创新的“菜中种群”策略，其中 来自许多不同神经典型和 16p11.2del 患者的干细胞系将被汇集到一个培养瓶中 询问表型差异，Aim 3 (R00) 将利用这些 scRNA-seq 技术来破译其作用。 16p11.2 基因在对神经发育很重要的特定通路中的成功完成。 可能导致治疗干预的遗传目标的识别，同时也显着改变 这些实验将提供新的领域对人类大脑疾病进行体外建模的方式。 为该提案的主要研究者 (PI) 提供有关 scRNA-seq 和生物信息学方法的培训，这些方法将服务于 作为一个独立研究实验室的基础，该实验室将使用干细胞衍生的神经细胞和大 转录组数据集与动物模型相结合，阐明细胞和分子机制 这项工作将在布罗德研究所和哈佛大学完成。 PI 将参加大学，那里有无数的技术和智力发展机会。 与导师和合作者定期举行会议，以获得有关实验设计和职业的反馈 PI 将参加哈佛大学的资助写作和项目管理课程，同时提高他的能力。 通过在国际科学会议上展示数据来培养沟通技巧 总的来说，这个职业。 发展计划将帮助PI在研究型学术机构建立自己的团队， 成为神经发育疾病建模领域的领导者。