Functional convergence following disruption of diverse genes associated with neurodevelopmental disorders

与神经发育障碍相关的多种基因被破坏后的功能趋同

基本信息

批准号：
10626945
负责人：
Kristen Jennifer Brennand
金额：
$ 75.78万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-20 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10626945
关键词：
16p11.2 Alleles Autopsy Biological Biological Assay Biology Chromatin Remodeling Factor Clustered Regularly Interspaced Short Palindromic Repeats Communication Communities Complex Data Data Set Development Disease Disparate Electronic Health Record Engineering Etiology Evaluation Female Gene Expression Gene Expression Profile Genes Genetic Genetic Transcription Genetic Variation Genetic study Genomics Glutamates Goals Heterozygote Human Individual Induced pluripotent stem cell derived neurons Link Methods Modeling Mutation National Institute of Mental Health Neurodevelopmental Deficit Neurodevelopmental Disorder Neurons Outcome Pathogenesis Pathway interactions Phenotype Predisposition Process Protein Truncation Proteins Relative Risks Research Resources Risk Risk Factors Sampling Series Synapses Testing Transcriptional Regulation autism spectrum disorder cell type chromatin modification chromatin remodeling data integration disorder risk druggable target fetal gene discovery genome-wide improved in silico induced pluripotent stem cell innovation insight loss of function loss of function mutation male neurodevelopment neuropsychiatric disorder novel parallelization postnatal risk variant stem cell model synaptic function therapeutic target transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY A complex interplay of genetic variation underlies predisposition for autism spectrum disorder (ASD). There is now strong evidence from large consortia studies that mutations in genes involved in chromatin modification, transcriptional regulation, and synaptic proteins confer substantial risk for ASD; however, the extent to which these genes are interconnected and ultimately converge on a small number of functional deficits is largely unknown. A critical need therefore exists to model new gene discoveries, to directly evaluate their functional impact, and to determine their points of convergence. Innovations from our team and others in high-throughput CRISPR-engineering have now made parallelized mechanistic studies tractable, and human induced pluripotent stem cell (hiPSCs) derived neurons are well-suited to test the impact of ASD risk variants predicted to exert their influence during fetal cortical development. Here, our multi-PI proposal will undertake an ambitious, systematic isogenic loss-of-function (LoF) mechanistic screen in a compendium of 48 of the most robust ASD risk genes discovered from the largest genetic studies to date. Moreover, our exciting preliminary results suggest that transcriptional signatures shared across neuronal models of ASD genes converge on critical regulatory nodes that result in synaptic deficits. Aim 1 will characterize isogenic glutamatergic and GABAergic neurons with highly penetrant LoF mutations in 48 genes associated with ASD risk at genome-wide significant thresholds and that are expressed in neurons. These analyses will identify transcriptional and functional signatures of individual ASD genes through RNAseq and a series of high-throughput phenotyping assays in both neuronal sub-types. Aim 2 will expand our Preliminary Results to discover convergent genes downstream of ASD risk loci, characterize the synaptic consequences of the ten most compelling discoveries from individual genes and/or convergent signatures, and integrate these data to explore the druggability of the convergent networks. Our overarching goal is to define any commonalities among diverse genes, pathways and networks that underlie ASD etiology, and to dramatically expand the list of possible therapeutic targets for ASD. These studies will generate an unprecedented isogenic resource of CRISPR-edited ASD genes, and matched RNAseq and cellular phenotyping in glutamatergic and GABAergic neurons, that will be provided for open distribution to the broader community through the NIMH RUDCR resource to yield new insights into neuropsychiatric disorders.

项目概要遗传变异的复杂相互作用是自闭症谱系障碍 (ASD) 易感性的基础。有现在来自大型联盟研究的有力证据表明，涉及染色质修饰的基因突变，转录调控和突触蛋白给 ASD 带来巨大风险；然而，在多大程度上这些基因是相互关联的，最终集中在少数功能缺陷上未知。因此，迫切需要对新基因发现进行建模，直接评估其功能影响，并确定其交汇点。我们团队和其他人在高通量方面的创新 CRISPR 工程现已使并行机制研究变得易于处理，并且人类诱导多能性干细胞 (hiPSC) 衍生的神经元非常适合测试 ASD 风险变异的影响，预计这些变异会发挥其作用影响胎儿皮质发育。在这里，我们的多 PI 提案将进行一项雄心勃勃的、系统的对 48 个最强大的 ASD 风险基因进行了等基因功能丧失 (LoF) 机制筛选迄今为止最大的遗传学研究发现的。此外，我们令人兴奋的初步结果表明自闭症谱系障碍（ASD）基因的神经元模型共享的转录特征汇聚在关键的调控节点上导致突触缺陷。目标 1 将表征同基因谷氨酸能和 GABA 能神经元，具有高度与 ASD 风险相关的 48 个基因的渗透性 LoF 突变处于全基因组显着阈值，并且在神经元中表达。这些分析将识别个体 ASD 的转录和功能特征通过 RNAseq 和一系列高通量表型分析对两种神经元亚型进行基因分析。目标2 将扩展我们的初步结果，以发现 ASD 风险位点下游的趋同基因，表征来自个体基因和/或趋同基因的十个最引人注目的发现的突触后果签名，并整合这些数据来探索融合网络的成药性。我们的首要任务目标是定义 ASD 病因的不同基因、途径和网络之间的共性，并大幅扩大自闭症谱系障碍（ASD）可能的治疗靶点清单。这些研究将产生前所未有的 CRISPR 编辑的 ASD 基因同基因资源，以及匹配的 RNAseq 和细胞表型分析谷氨酸能和 GABA 能神经元，将开放分发给更广泛的社区通过 NIMH RUDCR 资源，对神经精神疾病产生新的见解。