Genes and developmental signaling pathways in neuropsychiatric disorders

神经精神疾病的基因和发育信号通路

• 批准号: 9229296
• 负责人: Tracy L YOUNG-PEARSE
• 金额: $ 8.87万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9229296
• 关键词: 22q11.2; Address; Affect; Autopsy; Binding; Biological Assay; Brain; Cell Culture Techniques; Cell Line; Cell Proliferation; Cell model; Cells; Cerebral cortex; Collaborations; Collection; Complement; Complex; Copy Number Polymorphism; DISC1 gene; Data; Defect; Development; Developmental Gene; Disease; Electrophysiology (science); Electroporation; Embryo; Engineering; Equilibrium; Functional disorder; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Models; Genomics; Goals; Health; Human; Human Pathology; In Vitro; Individual; Lead; Letters; Link; Major Mental Illness; Masks; Mediating; Mental disorders; Methods; Microelectrodes; Molecular; Mutation; Neurites; Neuronal Differentiation; Neurons; Onset of illness; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Phosphotransferases; Play; Process; Protein Isoforms; Reelin Signaling Pathway; Research; Risk; Rodent; Rodent Model; Role; Schizophrenia; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Synapses; Time; Tissue Model; Transplantation; WNT Signaling Pathway; brain tissue; cell motility; gene transplantation for gene therapy; genetic linkage; genetic variant; human stem cells; human subject; in utero; in vivo; induced pluripotent stem cell; knock-down; migration; mouse model; mutant; neurodevelopment; neurogenesis; neuropsychiatric disorder; neurotransmission; overexpression; patient subsets; precursor cell; risk variant; signal processing; synaptogenesis; transcription activator-like effector nucleases

DESCRIPTION (provided by applicant): The development of the mammalian cortex is a complex process that requires the precise coordination of neuronal proliferation, migration, neurite outgrowth and synaptogenesis. Defects in corticogenesis are thought to underlie aspects of several neuropsychiatric disorders including schizophrenia (SCZ). In the past decade, several genes involved in neurodevelopment have been linked to SCZ and related disorders. A number of these genes act in either the canonical WNT or Reelin (RELN) signaling pathways, which have central roles in proliferation and migration, respectively. These pathways are traditionally thought to act independently at different times in the development of a neuron. However, shared molecular factors such as DISC1 exist between these pathways. Our overarching hypothesis is that the WNT and RELN pathways interact to regulate the developmental switch between proliferation and migration. Further, we posit that disruption of genes shared between these pathways, such as DISC1, lead to increased risk for psychiatric disorders. Our lab has established isogenic human induced pluripotent stem cells (iPSCs) that have a targeted mutation at the DISC1 locus near the site of a chr(1;11) balanced translocation linked to major mental disorders. We will use these lines to address if and how this disease-relevant mutation affects certain neurodevelopmental processes. Importantly, phenotypes observed will be examined in hiPSCs derived from human subjects with SCZ harboring the chr(1;11) translocation. Further, in order to examine whether the molecular and cellular phenotypes extend beyond DISC1 disruption, a limited analysis will be performed in hiPSCs from SCZ patients with CNVs linked to mental illness. To complement these in vitro studies, postmortem human brain tissue and rodent models will be utilized in parallel to establish the in vivo relevance of key findings. In Aim 1, we examine DISC1 isoform expression in isogenic and patient-derived wild-type and mutant lines over neuronal differentiation. We analyze the maturation of these neurons via gene expression profiling, morphometric analyses, and electrophysiological assays. In Aim 2, we investigate the WNT and RELN signal transduction pathways, and compare the impact of DISC1 disruption with SCZ-linked copy number variants (CNVs) on these pathways. Lastly, in Aim 3 we analyze the effects of DISC1 disruption and SCZ-linked CNVs on proliferation and migration in vivo in the embryonic rodent brain via the complementary methods of in utero electroporation to knock down or overexpress genes and transplantation of human NPCs harboring these disruptions. Taken together, we aim to identify how the effects of these mutations converge to result in altered cortical circuitry and the onset o mental illness. Under this research plan, we take initial steps to compare how specific genetic disruptions with strong effect on risk for mental illness affect developmental signaling pathways and processes in human neuronal cells.

描述（由申请人提供）：哺乳动物皮质的发展是一个复杂的过程，需要精确协调神经元增殖，迁移，神经突生长和突触发生。皮质生成中的缺陷被认为是包括精神分裂症（SCZ）在内的几种神经精神疾病的方面的基础。在过去的十年中，涉及神经发育的几个基因与SCZ和相关疾病有关。这些基因的许多作用在规范的Wnt或Reelin（Reln）信号通路中，它们分别在增殖和迁移中具有中心作用。传统上，这些途径在神经元的发展中在不同时期独立起作用。但是，这些途径之间存在共享的分子因素，例如二张圆盘。我们的总体假设是，Wnt和Reln途径相互作用以调节增殖和迁移之间的发育切换。此外，我们认为这些途径之间共享基因的破坏，例如DISC1，导致了精神疾病的风险增加。我们的实验室已经建立了同基因的人类诱导的多能干细胞（IPSC），这些干细胞（IPSC）在CHR（1; 11）位点附近的圆盘基因座的靶向突变，与主要精神障碍有关。我们将使用这些线路来解决该与疾病相关的突变如何影响某些神经发育过程。重要的是，观察到的表型将在源自人类受试者的HIPSC中检查，其中SCZ具有CHR（1; 11）易位。此外，为了检查分子和细胞表型是否超出了椎间盘的破坏，将在与精神疾病有关的CNV患者的HIPSC中进行有限的分析。为了补充这些体外研究，将同时利用死后人脑组织和啮齿动物模型来建立关键发现的体内相关性。在AIM 1中，我们检查了神经元分化的同基因和患者衍生的野生型和突变型线中的DISC1同工型表达。我们通过基因表达分析，形态分析和电生理测定法分析了这些神经元的成熟。在AIM 2中，我们研究了Wnt和RELN信号转导途径，并将盘式破坏与SCZ连接拷贝数变体（CNV）对这些途径的影响进行比较。最后，在AIM 3中，我们通过子宫内电穿孔中的互补方法分析了圆盘1中断和SCZ连接的CNV对胚胎啮齿动物大脑中体内增殖和迁移的影响，以敲低或过表达基因以及对携带这些破坏的人NPC的转移。综上所述，我们旨在确定这些突变的影响如何融合，从而导致皮质回路改变和发作O精神疾病。在该研究计划下，我们采取初步步骤来比较对精神疾病风险有强大影响的特定遗传破坏如何影响人类神经元细胞中的发育信号通路和过程。