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，残疾和强烈的遗传疾病。验尸研究表明减少 精神分裂症患者的皮质树突状脊柱密度，与结构神经成像一致 研究。同样，基因组数据将最大影响的精神分裂症相关的共同风险变异与 在啮齿动物模型中破坏修剪。这些收敛的证据表明，小胶质细胞介导 修剪异常可能是导致精神分裂症中观察到的神经病理学的原因 认识到选择性吞没小胶质细胞的突触的重要性 神经发育。但是，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中获得活细胞，以便快速筛查和 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 通过从患者血液隔离的单核细胞中重编程诱导的小胶质细胞进行修剪，并分析 它们具有从诱导的神经培养物中得出的孤立突触（突触体） 多能干细胞（IPSC）。在初步研究中，他们证明了 小胶质细胞和精神分裂症患者的突触体的异常，并救出 这种异常以小分子探针的剂量响应方式。 拟议的调查将使用非常大的患者来确认并扩展这些结果 细胞生物库由研究者开发。具体而言，这项研究将产生新的和充分的 表征了来自50名患者的诱导小胶质细胞培养物和IPSC衍生的神经培养 精神分裂症和50岁，性别和祖先匹配的健康对照。这些患者衍生的试剂将 用于检验小胶质细胞介导的突触修剪的功能差异。 患者和对照（AIM 1）。这些测定也将应用于屏幕小分子以识别 基于有希望的初步数据（AIM 2），基于突触修剪的其他调节剂。并联，高 吞吐量化学基因组方法将用于表征这些小的转录组效应 小胶质细胞上的分子心形扰，可深入了解作用机理并进一步促进 化学屏幕（AIM 3）。 这些研究将共同​​验证该平台的未来高通量筛查工作。 在新颖的治疗学上。该项目汇集了一个具有蜂窝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造关键资源，并具有高度 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。