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: 10614930
• 负责人: ROY H. Perlis
• 金额: $ 61.09万
• 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 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614930
• 关键词: Address; Adolescence; Adverse effects; Age; Age of Onset; Autopsy; Biochemical; Biological Assay; Biopsy; Blood; Blood specimen; Brain; Cell Reprogramming; 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; Human; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Investigation; Libraries; Link; Magnetic Resonance Imaging; Maintenance; Mediating; Mental disorders; Mentally Disabled Persons; Methods; Microglia; Minocycline; Modeling; Modeling of Functional Interactions; Neurobiology; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurons; Orthologous Gene; Pathogenesis; Patients; Phagocytosis; Prevalence; Rapid screening; Reagent; Research Personnel; Resources; Rodent; Rodent Model; Sampling; Schizophrenia; Serum; 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; neural; neurodevelopment; neuroimaging; neuropathology; novel; novel therapeutics; pharmacologic; relapse prevention; risk variant; screening; sex; small molecule; synaptic inhibition; 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的、致残的、具有强烈遗传性的疾病。验尸报告显示 精神分裂症患者皮质树突棘密度与结构神经影像学结果一致 问题研究同样，基因组数据将精神分裂症相关的最大影响的常见风险变异与 在啮齿动物模型中破坏修剪。这些证据表明，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， 公认的重要性，选择性吞噬突触的小胶质细胞作为修剪的手段，在正常 神经发育然而，大规模的人类小胶质细胞在疾病中的功能研究受到阻碍 由于难以从精神分裂症患者中获得适于快速筛查的活细胞， 量化功能评估。 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。 通过重编程从患者血液分离的单核细胞中修剪诱导的小胶质细胞，并测定 它们与来自神经培养物的分离的突触（突触体）一起从诱导的 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的异常， 用小分子探针以剂量响应的方式检测这些异常。 拟议的研究将使用一个非常大的患者来源的 研究人员开发的细胞生物库。具体而言，这项研究将产生新的和全面的 表征了来自50名患有神经胶质瘤的个体的诱导的小胶质细胞培养物和iPSC衍生的神经培养物。 精神分裂症和50名年龄、性别和血统匹配的健康对照。这些患者衍生试剂将 用于检测小胶质细胞介导的突触修剪的功能差异， 患者和对照组（目标1）。这些分析也将被应用于筛选小分子，以识别 突触修剪的其他调节剂，建立在有希望的初步数据（目标2）。平行，高 通量化学基因组学方法将被应用于表征这些小分子的转录组学效应。 分子干扰小胶质细胞，提供深入了解的作用机制，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未来高通量筛选工作的平台， 新疗法的研究该项目汇集了一个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。除了调查这些主要的 假设，该项目将为神经生物学社区创造一个重要的资源，具有高- 扩展患者和健康对照细胞的完全注释和可共享的生物库的维度数据。