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

• 批准号: 10407989
• 负责人: Kristen Jennifer Brennand
• 金额: $ 79.65万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/10407989
• 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 Discovery; Data Set; Development; Disease; Electronic Health Record; Engineering; Etiology; Evaluation; Female; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genetic Variation; Genetic study; Genomics; Glutamates; Goals; 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; Proteins; Relative Risks; Research; Resources; Risk; Risk Factors; Sampling; Series; Synapses; Testing; Transcriptional Regulation; autism spectrum disorder; cell type; chromatin modification; chromatin remodeling; 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; postnatal; risk variant; stem cell model; synaptic function; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics

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.

项目摘要 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，来自大型财团研究的有力证据表明，基因突变涉及染色质修饰， 转录调控和突触蛋白赋予ASD的实质性风险;然而， 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，迫切需要对新的基因发现进行建模，以直接评估其功能， 影响，并确定它们的汇合点。我们的团队和其他人在高通量方面的创新 CRISPR工程现在已经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于测试ASD风险变体的影响，这些变体被预测发挥其功能。 影响胎儿皮质发育。在这里，我们的多PI建议将进行一个雄心勃勃的，系统的 在48个最强大的ASD风险基因的纲要中进行同基因功能丧失（LoF）机制筛选 迄今为止最大的遗传学研究发现。此外，我们令人兴奋的初步结果表明， ASD基因的神经元模型之间共享的转录特征汇聚在关键调控节点上 导致突触缺陷目的1将表征具有高度同源性的谷氨酸能和GABA能神经元， 在全基因组显著阈值下，48个与ASD风险相关的基因中的渗透性LoF突变， 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在两种神经元亚型中检测基因。目的2 将扩大我们的初步结果，以发现ASD风险基因座下游的会聚基因， 突触后果的十大最引人注目的发现从个别基因和/或收敛 签名，并整合这些数据，以探索聚合网络的药物性。我们的总体 目的是确定ASD病因学基础的不同基因、途径和网络之间的任何共性， 并极大地扩展了ASD可能的治疗靶点。这些研究将产生一个 CRISPR编辑的ASD基因的前所未有的同基因资源，以及匹配的RNAseq和细胞表型 在多巴胺能和GABA能神经元中，将提供给更广泛的社区开放分配 通过NIMH RUDCR资源，对神经精神疾病产生新的见解。