Patient-derived Models of Synaptic Pruning in Schizophrenia

精神分裂症患者衍生的突触修剪模型

• 批准号: 9981011
• 负责人: ROY H. Perlis
• 金额: $ 64.3万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-19 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981011
• 关键词: Address; Adolescence; Adverse effects; Age; Age of Onset; Autopsy; Biochemical; Biological Assay; Biopsy; Blood; Blood specimen; Brain; Cell model; Cells; Cellular Assay; Chemicals; Chronic; Coculture Techniques; Code; Communities; Complement; Complex; Data; Dendritic Spines; Development; Disease; Dose; Economic Burden; Environment; Exhibits; Future; Genes; Genetic Diseases; Genome; Genomics; Heritability; Hippocampus (Brain); Human; In Vitro; Individual; Investigation; Libraries; Link; Magnetic Resonance Imaging; Maintenance; Mediating; Mental disorders; Methods; Microglia; Minocycline; Modeling; Modeling of Functional Interactions; Neurobiology; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurons; Orthologous Gene; Pathogenesis; Patients; Phagocytosis; Pharmacology; Prevalence; Reagent; Relapse; Research Personnel; Resources; Rodent; Rodent Model; Sampling; Schizophrenia; Serum; Structure; Synapses; Synaptosomes; Thick; Vertebral column; biobank; clinical phenotype; density; disability; effective therapy; emerging adult; functional restoration; genome wide association study; genomic data; high throughput screening; high-throughput drug screening; human data; imaging study; induced pluripotent stem cell; insight; monocyte; multidimensional data; neurodevelopment; neuroimaging; neuropathology; novel; novel therapeutics; prevent; relating to nervous system; risk variant; screening; sex; small molecule; synaptic pruning; transcriptome; transcriptomics

Schizophrenia is a chronic, disabling, and strongly heritable illness. Postmortem studies suggest reduced cortical dendritic spine density among schizophrenia patients, consistent with structural neuroimaging studies. Likewise, genomic data links schizophrenia-associated common risk variants of greatest effect to disrupted pruning in a rodent model. These convergent lines of evidence suggest that microglia-mediated pruning abnormalities may be responsible for the observed neuropathology in schizophrenia, extending the recognized importance of selective engulfment of synapses by microglia as a means of pruning in normal neurodevelopment. However, large-scale functional studies of human microglia in disease are hampered by difficulties in obtaining living cells from individuals with schizophrenia amenable to rapid screening and quantitative functional assessments. The investigators have recently developed and validated patient-specific models of microglia-mediated pruning by reprogramming induced microglial cells from patient blood isolated monocytes, and assaying them with isolated synapses (synaptosomes) derived from neural cultures differentiated from induced pluripotent stem cells (iPSCs). In preliminary studies, they have demonstrated robust evidence of abnormalities in both microglia as well as synaptosomes from individuals with schizophrenia, and rescued such abnormalities in a dose-responsive fashion with a small molecule probe. The proposed investigation will confirm and extend these results using a very large patient-derived cellular biobank developed by the investigators. Specifically, this study will generate new and fully characterized induced microglia cultures and iPSC-derived neural cultures from 50 individuals with schizophrenia and 50 age, sex, and ancestry-matched healthy controls. These patient-derived reagents will be utilized in an assay to examine functional differences in microglia-mediated synaptic pruning from patients and controls (Aim 1). These assays will also be applied to screen small molecules to identify additional modulators of synaptic pruning, building on promising preliminary data (Aim 2). In parallel, high throughput chemical genomic methods will be applied to characterize transcriptomic effects of these small molecule perturbagens on microglia, providing insight into mechanism of action and facilitating further chemical screens (Aim 3). Together, these studies will further validate the platform for future high-throughput screening efforts aimed at novel therapeutics. The project brings together a team with expertise in cellular modeling, transcriptomics, clinical phenotyping, and small molecule screening. Beyond investigating these principal hypotheses, the project will create a critical resource for the neurobiological community, with high- dimensionality data extending a fully annotated and shareable biobank of patient and healthy control cells.

精神分裂症是一种慢性、致残且具有强烈遗传性的疾病。尸检研究表明减少 精神分裂症患者的皮质树突棘密度，与结构神经影像一致 研究。同样，基因组数据将与精神分裂症相关的常见风险变异联系起来，对精神分裂症影响最大 啮齿动物模型中的修剪中断。这些汇聚的证据表明小胶质细胞介导的 修剪异常可能是精神分裂症中观察到的神经病理学的原因，延长了 认识到小胶质细胞选择性吞噬突触作为正常修剪手段的重要性 神经发育。然而，人类小胶质细胞在疾病中的大规模功能研究受到阻碍 由于难以从精神分裂症患者身上获得活细胞并进行快速筛选 定量功能评估。 研究人员最近开发并验证了小胶质细胞介导的患者特异性模型 通过从患者血液分离的单核细胞中重新编程诱导的小胶质细胞进行修剪，并进行测定 它们具有分离的突触（突触体），这些突触源自神经培养物，与诱导分化 多能干细胞（iPSC）。在初步研究中，他们已经证明了强有力的证据 精神分裂症患者的小胶质细胞和突触体均出现异常，并被拯救 使用小分子探针以剂量响应方式检测此类异常。 拟议的调查将使用非常大的源自患者的数据来确认并扩展这些结果 研究人员开发的细胞生物库。具体来说，这项研究将产生新的、全面的 对 50 名个体的诱导小胶质细胞培养物和 iPSC 衍生的神经培养物进行了表征 精神分裂症患者和 50 名年龄、性别和血统相匹配的健康对照者。这些源自患者的试剂将 可用于检测小胶质细胞介导的突触修剪的功能差异 患者和对照（目标 1）。这些测定还将用于筛选小分子以识别 基于有希望的初步数据（目标 2），突触修剪的额外调节剂。并联，高 通量化学基因组方法将用于表征这些小分子的转录组效应 分子扰动小胶质细胞，提供对作用机制的深入了解并促进进一步 化学筛选（目标 3）。 总之，这些研究将进一步验证该平台用于未来的高通量筛选工作，旨在 在新颖的疗法中。该项目汇集了一支具有细胞建模专业知识的团队， 转录组学、临床表型和小分子筛选。除了调查这些校长之外 假设，该项目将为神经生物学界创建一个重要的资源，具有高 维度数据扩展了患者和健康对照细胞的完全注释和可共享的生物库。