TAOK2 Kinase Signaling in Human Neural Stem Cell Development

人类神经干细胞发育中的 TAOK2 激酶信号转导

• 批准号: 10064147
• 负责人: Smita Yadav
• 金额: $ 44.99万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-02 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064147
• 关键词: 16p11.2; 3-Dimensional; Affect; Biochemical; Biological Assay; Biological Models; Biology; Brain; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Centrioles; Centrosome; Cilia; Clinical; Complex; Copy Number Polymorphism; Data; Defect; Development; Disease; Dissection; Event; FRAP1 gene; Fluorescence; Functional disorder; Future; Genes; Genetic; Growth; Human; Human Development; Human Engineering; Image; Knock-out; Length; Maps; Mass Spectrum Analysis; Mediating; Mental disorders; Methodology; Microscopy; Molecular; Mothers; Mutation; Neurodevelopmental Disorder; Neurons; Organoids; Outcome; Pathogenesis; Pathology; Pathway interactions; Patients; Phosphorylation; Phosphotransferases; Protein-Serine-Threonine Kinases; Proteins; Proteome; Proteomics; Research Proposals; Resolution; Role; Sampling; Signal Pathway; Signal Transduction; Stem Cell Development; Structural defect; Structure; Susceptibility Gene; Synapses; Techniques; Testing; Work; autism spectrum disorder; base; chemical genetics; cilium biogenesis; clinical Diagnosis; endophenotype; experimental study; gain of function mutation; genetic risk factor; genome editing; genomic locus; human disease; induced pluripotent stem cell; induced pluripotent stem cell technology; innovation; insight; loss of function; nerve stem cell; nervous system disorder; neurodevelopment; neurogenesis; neuron development; neuropathology; neuroregulation; novel therapeutic intervention; null mutation; patient variability; sensor; stem cell model; stem cell proliferation; stem cell technology

Increasing evidence indicates that aberrations in neural stem cell proliferation and early neurogenesis are critically involved in the pathogenesis of neurodevelopmental and psychiatric disorders. Despite being implicated in pathophysiology of several neurological diseases, the mechanisms through which human kinome controls neurogenesis and how its dysfunction manifests in disease remain major gaps in the field of neurodevelopmental biology. In this proposal, we will investigate the role of an autism susceptibility gene, TAOK2, which encodes a serine threonine kinase, in human neural stem cell development and function. Both loss-of-function and activating TAOK2 mutations have been associated with autism spectrum disorders (ASD). Further, TAOK2 is one of the genes in the 16p11.2 genomic locus, copy number variation (CNV) of which is the most prevalent genetic risk factor associated with ASD. While work by us and others has demonstrated compelling evidence that TAOK2 is important for neuronal and synaptic development, the role of TAOK2 in human neural progenitor cell (NPC) development and differentiation has not been investigated. Further, the contribution of TAOK2 in the pathology associated with 16p11.2 copy number variation is unknown. Based on the strength of our preliminary findings, the central hypothesis of this research proposal is that TAOK2 kinase orchestrates a signaling hub at the centrosome that regulates human neural progenitor stem cell development, and that perturbation of this signaling pathway contributes to pathogenesis of ASD. We will elucidate the role of TAOK2 in NPC development through use of 2D and 3D human induced pluripotent stem cell (hiPSC) models (Aim1). Next, mechanisms through which TAOK2 regulates ciliary growth and signaling will be determined using super-resolution imaging and biochemical approaches (Aim2). The contribution of TAOK2 in the ciliary defects observed in patient-derived NPCs from 16p11.2 deletion and duplication carriers will be determined through application of quantitative proteomics and genome editing techniques (Aim3). Utilizing a combination of innovative approaches and human disease relevant model systems, we seek to understand how TAOK2 kinase signaling mediates neural stem cell development, and how perturbations in its signaling pathways contribute to the clinical neuropathology of 16p11.2 CNV. !

越来越多的证据表明神经干细胞增殖和早期神经发生的畸变 与神经发育和精神疾病的发病机制密切相关。尽管是 与多种神经系统疾病的病理生理学有关，人类激酶组的机制 控制神经发生及其功能障碍如何在疾病中表现仍然是该领域的主要空白 神经发育生物学。在这项提案中，我们将研究自闭症易感基因的作用， TAOK2 编码丝氨酸苏氨酸激酶，参与人类神经干细胞的发育和功能。两个都 功能丧失和激活 TAOK2 突变与自闭症谱系障碍 (ASD) 相关。 此外，TAOK2是16p11.2基因组位点中的基因之一，其拷贝数变异(CNV)为 与 ASD 相关的最普遍的遗传风险因素。虽然我们和其他人的工作已经证明 令人信服的证据表明 TAOK2 对神经元和突触发育很重要，TAOK2 在 人类神经祖细胞（NPC）的发育和分化尚未得到研究。此外， TAOK2 在与 16p11.2 拷贝数变异相关的病理学中的作用尚不清楚。基于 根据我们初步研究结果的强度，本研究提案的中心假设是 TAOK2 激酶 在中心体协调一个信号枢纽，调节人类神经祖细胞干细胞的发育， 该信号通路的扰动会导致自闭症谱系障碍 (ASD) 的发病机制。我们将阐明的作用 通过使用 2D 和 3D 人类诱导多能干细胞 (hiPSC) 模型，TAOK2 在 NPC 发育中的作用 （目标1）。接下来，将确定 TAOK2 调节纤毛生长和信号传导的机制 使用超分辨率成像和生化方法（Aim2）。 TAOK2对睫状体的贡献 将确定来自 16p11.2 缺失和重复携带者的患者来源的 NPC 中观察到的缺陷 通过应用定量蛋白质组学和基因组编辑技术（Aim3）。使用组合 在创新方法和人类疾病相关模型系统的研究中，我们试图了解 TAOK2 如何 激酶信号传导介导神经干细胞发育，以及其信号通路的扰动如何 有助于 16p11.2 CNV 的临床神经病理学。 ！