High-throughput in vivo and in vitro functional and multi-omics screens of neuropsychiatric and neurodevelopmental disorder risk genes

神经精神和神经发育障碍风险基因的高通量体内和体外功能和多组学筛选

• 批准号: 10643398
• 负责人: Kristen Jennifer Brennand
• 金额: $ 112.66万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643398
• 关键词: 3-Dimensional; Affect; Arousal; Behavior; Behavioral; Biological; Biological Assay; Biology; Brain; CRISPR screen; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Coupled; Data; Databases; Development; Disease; Estrogens; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Genomics; Goals; Human; Human Engineering; In Vitro; Induced pluripotent stem cell derived neurons; Link; Maps; Mental disorders; Methods; Molecular; Neurodevelopmental Disorder; Neuroglia; Neurons; Nuclear Pore Complex; Optics; Organoids; Outcome; Pathway interactions; Phenotype; Resources; Role; Sensory; Series; Structure; System; Technology; Time; Zebrafish; behavioral pharmacology; behavioral phenotyping; brain cell; cell type; data integration; epigenomics; gene discovery; gene function; genomic data; in vivo; induced pluripotent stem cell; innovation; insight; loss of function; multiple omics; mutant; neurodevelopment; neuropsychiatric disorder; novel; risk variant; screening; single cell sequencing; single-cell RNA sequencing; tool; tool development; transcriptomics

PROJECT SUMMARY There is a critical need to develop high-throughput scalable assays to identify biological mechanisms underlying risk genes in neurodevelopmental and neuropsychiatric disorders (NPD). In this proposal, we aim to leverage the unique advantages of two scalable systems – human induced pluripotent stem cells (hiPSCs) and zebrafish – to perform parallel functional assays of NPD genes in vitro and in vivo, and to pilot the development of innovative spatial multi-omics technologies applicable across systems. We propose to establish an Assay and Data Generation Center (ADGC) as part of the SSPsyGene Consortium that capitalizes on the unique and complementary expertise of our labs in large-scale hiPSC CRISPR screens (Brennand), high-throughput zebrafish screens (Hoffman), and cutting-edge multi-omics tool development (Fan). Our goal is to gain novel insights into the convergent and divergent mechanisms by which diverse NPD gene loss of function affects neurodevelopment at the molecular, cellular, structural, circuit, and behavioral levels. We propose to screen 250 NPD genes using a tiered strategy in hiPSCs and zebrafish by conducting pooled and arrayed transcriptomic and phenotypic screens in hiPSCs-derived neurons and glia (Aim 1), CRISPR screens in zebrafish to assess the effects of gene loss of function on whole-brain structure, activity, and basic behaviors (Aim 2), and spatial transcriptomic and multi-omic CRISPR screens to investigate the transcriptional effects of NPD gene disruption in both systems (Aim 3). We will advance the field by identifying biologically relevant phenotypes resulting from NPD gene loss of function across multiple scales, informing gene prioritization schema, and establishing new spatial multi-omics platforms for the functional analysis of NPD genes. These studies will generate an unprecedented resource of matched molecular, cellular, structural, circuit, and behavioral data in hiPSCs and zebrafish, which will be provided for open distribution to the broader community to yield new insights into NPD.

项目概要 迫切需要开发高通量可扩展的检测方法来识别潜在的生物机制 在本提案中，我们的目标是利用神经发育和神经精神疾病（NPD）的风险基因。 两种可扩展系统的独特优势——人类诱导多能干细胞 (hiPSC) 和斑马鱼 – 在体外和体内对 NPD 基因进行并行功能测定，并试点开发 我们建议建立一种适用于各个系统的创新空间多组学技术。 数据生成中心 (ADGC) 作为 SSPsyGene 联盟的一部分，利用独特和 我们实验室在大规模 hiPSC CRISPR 筛选 (Brennand)、高通量方面的专业知识互补 斑马鱼筛选（霍夫曼）和尖端多组学工具开发（范）我们的目标是获得新颖。 深入了解不同 NPD 基因功能丧失的趋同和趋异机制 我们提出，在分子、细胞、结构、回路和行为水平上影响神经发育。 通过进行汇集和阵列，在 hiPSC 和斑马鱼中使用分层策略筛选 250 个 NPD 基因 hiPSC 衍生的神经元和神经胶质细胞的转录组和表型筛选（目标 1），CRISPR 筛选 斑马鱼评估基因功能丧失对全脑结构、活动和基本行为的影响 （目标 2），以及空间转录组和多组学 CRISPR 筛选，以研究转录效应 两个系统中的 NPD 基因破坏（目标 3）我们将通过识别生物学相关性来推进该领域的发展。 NPD 基因在多个尺度上功能丧失导致的表型，为基因优先排序提供信息 模式，并建立新的空间多组学平台用于 NPD 基因的功能分析。 研究将产生前所未有的匹配分子、细胞、结构、电路和 hiPSC 和斑马鱼的行为数据，将向更广泛的社区公开分发 产生对 NPD 的新见解。