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: 10334934
• 负责人: Michael Frederick Wells
• 金额: $ 2.26万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10334934
• 关键词: 16p11.2; Affect; Agonist; 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 Sequence Alteration; 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; Genetic Transcription; 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; 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; Series; Signal Pathway; Susceptibility Gene; Syndrome; System; Techniques; Technology; Therapeutic Intervention; 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; repetitive behavior; response; single-cell RNA sequencing; skills; social; stem cell model; 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.

项目总结/文摘