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

项目摘要 迫切需要开发高通量可扩展的检测方法，以确定潜在的生物学机制。 神经发育和神经精神障碍（NPD）的风险基因。在这份提案中，我们旨在利用 两个可扩展系统的独特优势-人类诱导多能干细胞（hiPSC）和斑马鱼 - 在体外和体内进行NPD基因的平行功能测定，并试点开发 创新的空间多组学技术适用于不同系统。我们建议建立一个分析， 数据生成中心（ADGC）作为SSPsyGene联盟的一部分， 我们的实验室在大规模hiPSC CRISPR筛选（Brennessee），高通量， 斑马鱼筛选（霍夫曼）和尖端的多组学工具开发（范）。我们的目标是获得新的 深入了解不同NPD基因功能丧失的会聚和发散机制 在分子、细胞、结构、电路和行为水平上影响神经发育。我们提出 在hiPSC和斑马鱼中使用分层策略筛选250个NPD基因， 在hiPSC衍生的神经元和神经胶质细胞中进行转录组学和表型筛选（Aim 1）， 斑马鱼，以评估基因功能丧失对全脑结构、活动和基本行为的影响 (Aim 2），以及空间转录组学和多组学CRISPR筛选，以研究 两种系统中的NPD基因破坏（目的3）。我们将通过识别生物相关的 NPD基因功能丧失导致的表型跨越多个尺度，告知基因优先级 模式，并建立新的空间多组学平台的NPD基因的功能分析。这些 研究将产生前所未有的匹配分子，细胞，结构，电路， hiPSC和斑马鱼的行为数据，将提供给更广泛的社区开放分发 对NPD产生新的见解。

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.