UCLA High-Throughput Neuropsychiatric Disorder Phenotyping Center (UCLA HT-NPC)

加州大学洛杉矶分校高通量神经精神疾病表型中心 (UCLA HT-NPC)

基本信息

批准号：
10643541
负责人：
DANIEL H GESCHWIND
金额：
$ 165.22万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
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/10643541
关键词：
3-Dimensional Address Alleles Anatomy Architecture Astrocytes Biological Biological Assay Biological Testing Brain CRISPR/Cas technology Calcium Cell Line Cell Survival Cells Cerebrum Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Communities Core Facility Coupled Custom Data Defect Development Developmental Delay Disorders Dideoxy Chain Termination DNA Sequencing Disease Disease Pathway Essential Genes Female Gene Expression Genes Genetic Heterogeneity Genetic study Genome engineering Human Human Genetics Image In Vitro Intellectual functioning disability Knock-out Link Measures Methods Modeling Molecular Morphology Mutation Nervous System Neurobiology Neurodevelopmental Disorder Neurons Neurophysiology - biologic function Nuclear RNA Optics Organoids Pathway interactions Phenotype Physiological Process Production Proliferating Proteins Reproducibility Research Personnel Resolution Resource Sharing Role Schizophrenia Small Nuclear RNA Synapses System Testing Time autism spectrum disorder cell type data sharing density disease phenotype disorder risk engineered stem cells excitatory neuron gene function genetic manipulation genetic risk factor high dimensionality human disease human embryonic stem cell human model human pluripotent stem cell human stem cells in vivo induced pluripotent stem cell inhibitory neuron knockout gene male model organism molecular phenotype multimodality nerve stem cell neural neurite growth neurodevelopment neurogenesis neuronal cell body neuropsychiatric disorder novel null mutation phenotypic data risk variant screening sensor sex single nucleus RNA-sequencing single-cell RNA sequencing small hairpin RNA stem cell model stem cells three dimensional cell culture three-dimensional modeling transcriptome sequencing voltage

项目摘要

Project Summary/Abstract Human genetic studies have identified hundreds of genes contributing to Neuropsychiatric and Neurodevelop- mental Disease (NPD) risk. But for most genes, their normal function or the consequences of their absence or reduction on neurodevelopment and neural function are not known. Here, we propose to address the substantial challenges of discerning potential functions of hundreds of NPD genes through the development of a High Throughput Neuropsychiatric Disease Phenotyping Center (UCLA HT-NPC), driven by the activity of 9 highly collaborative investigators (Aharoni, Bhaduri, Damoiseaux, Geschwind, Golshani, Kitai, Luo, Novich, and Wells) and two substantial core facilities (UCLA Molecular Screening Shared Resource and the Human Stem Cell and Genome Engineering Center). Through a tiered approach, we combine high throughput and high value, quantitative phenotyping with stem cell engineering to characterize the functional consequences of NPD gene knockouts (null alleles), a key initial step that will inform our understanding of disease pathways. In the first step, we will rapidly generate null alleles for 250 genes chosen by the Consortium using a rapid, high throughput lentiviral based system in hESCs. Viability and neural induction potential will be assessed, and quantitative phenotyping conducted using RNA-seq on all lines. Those genes passing viability and neural induction tests will be used in the production of clonal null hiPSC lines (male and female) for downstream phenotyping and wider distribution to the community. Subsequently, we will perform high throughput, quantitative, multi-scale phenotyping at the molecular, morphological, and physiological levels in both 2D and 3D hiPSC-based models of human cortical development. We leverage the relative strengths and scalability of each model to enable us to perform both snRNA and bulk RNA-seq, measure the maturation, morphology, and synaptic density of neural cells using automated imaging, including the multiplexed, protein-based CODEX (Phenocycler) platform, and characterize neuronal activity and synchronization through optical recordings using custom-built mini-scope arrays (STIMscope). By using multiple systems (e.g. hESC/hiPSC; gene editing, 2D and 3D cultures), we test biological reproducibility across systems and technical reproducibility through replication. The use of experimentally validated, quantitative phenotypes across multiple scales of analysis facilitates data sharing and comparisons with other SSPsyGene investigators and provides a template for the field more broadly.

项目概要/摘要人类遗传学研究已经确定了数百个与神经精神和神经发育有关的基因精神疾病（NPD）风险。但对于大多数基因来说，它们的正常功能或它们缺失或缺失的后果对神经发育和神经功能的减少尚不清楚。在此，我们建议解决实质性问题通过开发高水平的方法来识别数百个 NPD 基因的潜在功能的挑战吞吐量神经精神疾病表型中心 (UCLA HT-NPC)，由 9 个高度活跃的活动驱动合作研究人员（Aharoni、Bhaduri、Damoiseaux、Geschwind、Golshani、Kitai、Luo、Novich 和 Wells）和两个重要的核心设施（加州大学洛杉矶分校分子筛选共享资源和人类干细胞和基因组工程中心）。通过分层方法，我们将高吞吐量和高价值结合起来，通过干细胞工程进行定量表型分析来表征 NPD 基因的功能后果敲除（无效等位基因），这是一个关键的初始步骤，它将帮助我们了解疾病途径。在第一步中，我们将使用快速、高通量的方法快速为联盟选择的 250 个基因生成无效等位基因 hESC 中基于慢病毒的系统。将评估活力和神经诱导潜力，并定量使用 RNA-seq 对所有品系进行表型分析。那些通过活力和神经诱导测试的基因将用于生产无克隆 hiPSC 系（雄性和雌性），用于下游表型分析和更广泛的研究分发给社区。后续我们将进行高通量、定量、多尺度在基于 hiPSC 的 2D 和 3D 模型中进行分子、形态和生理水平的表型分析人类皮质发育。我们利用每个模型的相对优势和可扩展性，使我们能够执行 snRNA 和bulk RNA-seq，测量神经的成熟度、形态和突触密度使用自动成像的细胞，包括多重、基于蛋白质的 CODEX (Phenocycler) 平台，以及使用定制的微型显微镜通过光学记录来表征神经元活动和同步阵列（STIMscope）。通过使用多个系统（例如 hESC/hiPSC；基因编辑、2D 和 3D 培养），我们测试跨系统的生物再现性和通过复制的技术再现性。使用经过实验验证的跨多个分析尺度的定量表型有助于数据共享和与其他 SSPsyGene 研究人员进行比较，并为该领域提供更广泛的模板。