Pooled Optical Imaging, Neurite Tracing, and Morphometry Across Perturbations (POINT-MAP).

混合光学成像、神经突追踪和扰动形态测量 (POINT-MAP)。

• 批准号: 10741188
• 负责人: Joseph D. Buxbaum
• 金额: $ 46.48万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-03 至 2025-08-02
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10741188
• 关键词: Affect; Animal Model; Attention deficit hyperactivity disorder; Bar Codes; Benchmarking; Biological; Biological Models; Brain Diseases; Cell model; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive; Cognitive deficits; Complex; Development; Disease; Evaluation; Gene Cluster; Genes; Genetic; Genetic study; Genome; Genomic approach; Goals; Human; Image; Immunofluorescence Immunologic; Induced pluripotent stem cell derived neurons; Intellectual functioning disability; Investigation; Link; Measures; Methods; Microscopic; Molecular; Molecular Profiling; Morphology; Mutation; Neurites; Neurobiology; Neurodevelopmental Disorder; Neurons; Optics; Outcome; Pathogenesis; Pathway interactions; Periodicity; Phenotype; Recovery; Research; Retrieval; Risk; Role; Synapses; Technology; Therapeutic Intervention; Time; Validation; Variant; analysis pipeline; autism spectrum disorder; automated image analysis; disability; exome; functional genomics; genetic manipulation; genomic platform; high throughput screening; innovation; insight; knock-down; molecular phenotype; morphometry; new technology; novel; open source; optical imaging; precision medicine; reconstruction; risk variant; social deficits; success; synaptic function; synaptogenesis; targeted treatment; therapeutic target; transmission process

Project Summary Large scale exome-wide association studies have confidently implicated over 300 genes harboring rare mutations in the risk of neurodevelopmental disorders (NDDs), yet the specific molecular, cellular, and functional impacts of each are mostly unknown. Our long-term goal is to implement high-throughput functional genomics approaches to elucidate mechanisms for these genes and their variants. Neurons extend long and elaborately branched projections and connect to one another at synapses along these projections. A wealth of studies has implicated altered neuronal morphology and synapses in NDDs and other complex brain disorders. The overall objective of the proposed research is to combine a powerful human neuronal perturbation platform with an innovative cellular barcoding strategy to develop a novel technology enabling high throughput evaluation of the contributions of NDD risk genes on neuronal morphology and synaptogenesis. Our central hypothesis is that many NDD risk genes converge at the dysregulation of neuronal and synaptic function, and that we can identify such aberrations by measuring neuronal morphology after genetically perturbing risk genes. We aim to develop a novel scalable platform for Pooled Optical Imaging, Neurite Tracing, and Morphometry Across Perturbations (POINT-MAP). To do so, we will develop a novel optical cellular barcode that is linked to CRISPR perturbations, is neuronal cell-filling, and is identifiable by immunofluorescence. We will establish an automated image-analysis pipeline for barcode retrieval, neuronal tracing, and analysis of neuronal morphology and synaptic markers. We will demonstrate POINT-MAP in human induced pluripotent stem cell derived neurons to simultaneously assess the functional impact of 40 top NDD risk genes on neuronal and synaptic morphology.

项目概要 大规模全外显子组关联研究已确信涉及 300 多个基因 存在神经发育障碍（NDD）风险的罕见突变，但具体的 它们各自的分子、细胞和功能影响大多是未知的。我们的长期目标是 实施高通量功能基因组学方法来阐明这些机制 基因及其变体。神经元延伸出长且精心分支的投影并连接 沿着这些投影的突触处相互连接。大量研究表明改变 NDD 和其他复杂脑部疾病中的神经元形态和突触。整体 拟议研究的目标是结合强大的人类神经元扰动平台 采用创新的蜂窝条形码策略来开发新技术，实现高 NDD风险基因对神经元形态和功能的贡献的通量评估 突触发生。我们的中心假设是，许多 NDD 风险基因集中在 神经元和突触功能失调，我们可以通过以下方式识别这种异常 在遗传干扰风险基因后测量神经元形态。我们的目标是开发一个 用于混合光学成像、神经突追踪和形态测量的新型可扩展平台 扰动（点图）。为此，我们将开发一种新型光学蜂窝条形码 与 CRISPR 扰动有关，充满神经元细胞，并且可通过免疫荧光识别。 我们将建立一个自动化图像分析管道，用于条形码检索、神经元追踪、 以及神经元形态和突触标记的分析。我们将在以下位置演示 POINT-MAP 人类诱导多能干细胞衍生的神经元，同时评估功能 40 个 NDD 主要风险基因对神经元和突触形态的影响。