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: 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
• 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

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）的倾向。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， 转录调节，突触蛋白给ASD带来很大的风险；但是，在多大程度上 这些基因是相互联系的，最终在少数功能缺陷上收敛很大 未知。因此，存在着建模新基因发现的关键需求，直接评估其功能 影响并确定它们的收敛点。我们团队和其他人的创新 现在，CRISPR工程已经使平行的机械研究可进行，人类引起的多能 干细胞（HIPSC）衍生的神经元非常适合测试ASD风险变体的影响 胎儿皮质发育过程中的影响。在这里，我们的Multi-Pi提案将进行雄心勃勃的系统性 在48个最强大的ASD风险基因的纲要中，等源性丧失功能（LOF）机械屏幕 从迄今为止最大的遗传研究中发现。此外，我们令人兴奋的初步结果表明 跨ASD基因神经元模型共享的转录特征在关键调节节点上收敛 这导致突触缺陷。 AIM 1将表征具有高度的等源性谷氨酸能和GABA能神经元 在基因组明显阈值下与ASD风险相关的48个基因中的渗透力LOF突变，并且 用神经元表达。这些分析将确定单个ASD的转录和功能签名 通过RNASEQ的基因和两个神经元亚型中的一系列高通量表型测定。目标2 将扩大我们的初步结果，以发现ASD风险基因座下游的收敛基因，以表征 从单个基因和/或收敛的十个最引人注目的发现的突触后果 签名，并整合这些数据以探索收敛网络的可药用性。我们的总体 目标是定义ASD病因基础的不同基因，途径和网络之间的任何共同点， 并大大扩大ASD可能的治疗靶标的列表。这些研究将产生 CRISPR编辑的ASD基因的前所未有的ISEOGON资源，并匹配RNASEQ和细胞表型 在谷氨酸能和GABA能神经元中，将提供向更广泛的社区开放的 通过NIMH RUDCR资源，对神经精神疾病产生新的见解。