Functional analysis of autism risk genes during neural development using single cell seq

使用单细胞测序对神经发育过程中自闭症风险基因进行功能分析

• 批准号: 10334483
• 负责人: Antonio J Giraldez
• 金额: $ 56.75万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10334483
• 关键词: ASH1L gene; Address; Affect; Automobile Driving; Brain; CNTNAP1 gene; Cell Culture Techniques; Cell physiology; Cells; Chromatin; Chromatin Remodeling Factor; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Complex; Computing Methodologies; Development; Disease; Etiology; Exhibits; Gene Expression; Generations; Genes; Genetic; Genetic Models; Genetic Transcription; Genomics; Glutamates; Goals; Human; Individual; Knowledge; Life; Maps; Methods; Modeling; Modification; Molecular; Mutation; Neurons; Neurophysiology - biologic function; Organ; Organoids; Pathway interactions; Regulator Genes; Research; Resolution; Role; Shapes; Specific qualifier value; Synaptic plasticity; System; Testing; Work; Zebrafish; autism spectrum disorder; brain cell; cell type; combinatorial; disorder risk; estrogenic; experimental study; gene regulatory network; genome-wide; in vivo; innovation; insight; loss of function; loss of function mutation; mutant; nerve stem cell; neurodevelopment; neurogenesis; neuromechanism; novel; programs; protein protein interaction; relating to nervous system; response; risk variant; single cell sequencing; single-cell RNA sequencing; transcription factor; transcriptomics; vertebrate embryos

The brain represents a complex organ with a myriad of functionally diverse cell types. Autism spectrum disorder (ASD) is thought to alter the combinatorial transcriptional and post-transcriptional code responsible for specifying and maintaining different cell functions in the vertebrate brain. The proposed studies will test the central hypothesis that chromatin/transcriptional regulators identified as ASD risk genes act as developmental switches during early neurodevelopment, altering the gene expression network and subsequent cell function. In this proposal we combine single cell sequencing, with genetic, genomic and computational approaches in zebrafish and embryoid bodies to i) characterize the role of ASD-risk genes in healthy neural gene regulatory networks (Aim 1); 2) Interrogate the effects of ASD loss-of-function (LOF) mutations in different neural lineages (Aim 2). Cell types and developmental lineages within the brain that are more significantly affected by each candidate ASD risk gene will be identified using zebrafish and human cell–derived organoids. The experiments outlined in this proposal have the ultimate goal of identifying how ADS risk genes affect neural gene expression and cell function during vertebrate development.

大脑代表着一个复杂的器官，具有无数不同的功能 单元类型。自闭症谱系障碍(ASD)被认为改变了 转录和转录后代码负责指定和 维持脊椎动物大脑中不同的细胞功能。建议进行的研究 将检验染色质/转录调控因子识别出 因为ASD风险基因在早期起着发育开关的作用 神经发育，改变基因表达网络和后续细胞 功能。 在这项建议中，我们将单细胞测序与遗传、基因组 以及在斑马鱼和类胚体中的计算方法) 描述ASD危险基因在健康神经基因调控中的作用 网络(目标1)；2)询问ASD功能丧失(LOF)的影响 不同神经谱系的突变(目标2)。细胞类型与发育 大脑中受每个候选人影响更大的血统 将利用斑马鱼和人类细胞来源的细胞鉴定ASD风险基因 有机化合物。 这项提案中概述的实验的最终目标是 确定ADS风险基因如何影响神经基因表达和细胞功能 在脊椎动物的发育过程中。