Role of a psychiatric disease risk factor in synaptic function and gene transcription regulation

项目3：精神疾病危险因素在突触功能和基因转录中的作用 调控 摘要 遗传的复杂性是绝大多数精神障碍的基础，这使得对这些疾病的研究 非常具有挑战性。许多与风险相关的基因已被确定，但其生物学作用在很大程度上是 未知神经发育失调与结构和功能连接的改变被认为是 是许多神经精神疾病的基础，“突触疾病”是生物学的主要假设。 精神分裂症和其他主要精神疾病的基础。然而，对脑梗死的病理生理学知之甚少。 患者神经元中的突触，潜在的分子和细胞机制，以及精神疾病的程度 疾病可能共享这些机制。Disrupted in Schizophrenia 1（DISC 1）是一种基因， 与精神分裂症、双相情感障碍和其他主要精神疾病的风险增加有关。 紊乱大量的动物研究表明，DISC 1影响多种神经发育， 过程，包括突触的形成。理解重大精神疾病中的突触功能障碍 需要直接研究来自这些患者的人类神经元的突触特性， 紊乱重编程患者体细胞能够重演正常和病理人类 在确定的条件下的组织发育特性和识别细胞过程的新方法 潜在的复杂的人类疾病，这可能导致基于机制的药物发现。一种罕见的 发现DISC 1的C-末端的4个碱基对移码缺失与主要的 一个较小的美国家庭中的精神疾病（谱系H）。目前的项目是建立在我们最初的 结果显示，来自具有DISC 1突变的iPSC的前脑神经元表现出显著的突触功能。 缺陷和RNA-seq分析显示，大量神经元相关基因的显著失调， 神经元突触功能和精神疾病。使用来自特发性骨髓瘤的iPSC的最新研究 精神分裂症患者也表现出类似的突触功能缺陷，并共享多个失调基因。 项目3将检验一个假设，即精神障碍风险基因调节突触功能， 人类神经元通过生化和转录失调，一个核心缺陷，也可能是 存在于特发性精神分裂症和双相患者源性神经元中。目标1将表征细胞 H家系和特发性精神分裂症患者皮层神经元的分化表型 iPSCs。目的2将确定突变DISC 1在iPSC衍生的星形胶质细胞中的作用。目标3将评估神经元 突变DISC 1的亚型特异性对神经元发育、突触功能和转录的影响。每个 目标要求至少有一个学术和一个工业合作伙伴以及杨森的核心B参与 它被设计为在实验室之间进行交叉验证，并建立可以传播的协议， 进行进一步验证。细胞表型将与NCRCRG中的其他iPSC系进行比较，并将在NCRCRG中进行分析。 通过SBMRI的Core C进一步开发为小型化检测，用于未来的药物筛选。

Project 3: Role of a psychiatric disease risk factor in synaptic function and gene transcription regulation ABSTRACT Genetic complexity underlying the vast majority of mental disorders has made the study of these diseases exceptionally challenging. Many risk-associated genes have been identified but the biological role is largely unknown. Dysregulated neurodevelopment with altered structural and functional connectivity is believed to underlie many neuropsychiatric disorders and “a disease of synapses” is the major hypothesis for the biological basis of schizophrenia and other major psychiatric disorders. However, little is known about pathophysiology of synapses in patient neurons, underlying molecular and cellular mechanisms, and to what extent psychiatric disorders may share these mechanisms. Disrupted in Schizophrenia 1 (DISC1) is a gene in which mutations have been associated with increased risk for schizophrenia, bipolar disorder, and other major psychiatric disorders. A large number of animal studies have shown that DISC1 affects multiple neurodevelopmental processes, including synapse formation. Understanding synaptic dysfunction in major psychiatric disease requires direct investigation of synapse properties in human neurons derived from patients with these disorders. Reprogramming patient somatic cells enables recapitulation of normal and pathological human tissue developmental properties in defined conditions and a new way to identify the cellular processes underlying complex human diseases, which can lead to mechanism-based drug discovery. A rare mutation of a 4 base-pair frame-shift deletion at the C-terminus of DISC1 was discovered to co-segregate with major psychiatric disorders in a smaller American family (Pedigree H). The current project is built upon our initial results showing that forebrain neurons derived from iPSCs with the DISC1 mutation exhibit significant synaptic defects and RNA-seq analysis showed significant dysregulation of a large number of neuronal genes related to synaptic function and psychiatric disorders in patient neurons. Recent studies using iPSCs from idiopathic schizophrenia patients also showed similar defects in synaptic function and share multiple dysregulated genes. Project 3 will test the hypothesis that a psychiatric disorder risk gene modulates synaptic function of human neurons via biochemical and transcriptional dysregulation, a core defect that may also be present in idiopathic schizophrenia and bipolar patient-derived neurons. Aim 1 will characterize cellular phenotypes of human cortical neurons differentiated from Pedigree H and idiopathic schizophrenia patient iPSCs. Aim 2 will determine the role of mutant DISC1 in iPSC-derived astrocytes. Aim 3 will evaluate neuronal subtype specificity of mutant DISC1 effects on neuronal development, synaptic function and transcription. Each aim requires the involvement of at least one academic and one industrial partner as well as Core B at Janssen and is designed to incorporate cross-validation across labs and establish protocols that can be disseminated for further validation. Cellular phenotypes will be compared to other iPSC lines in the NCRCRG and will be further developed into miniaturized assays via Core C at SBMRI for future drug screens.