Neurogenesis: Career Development Plan in the Genetic and Modeling of Microcephaly

神经发生：小头畸形遗传和建模的职业发展计划

• 批准号: 8677906
• 负责人: Stephanie Lee Bielas
• 金额: $ 20.78万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-10 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8677906
• 关键词: Address; Affect; Agreement; Alleles; Animal Model; Animals; Antibodies; Brain; Cell Line; Cell Nucleus; Cell model; Cells; Cerebral cortex; Complement; Data; Defect; Development; Development Plans; Disease; Disease model; Evolution; Family; Fibroblasts; Foundations; Gene Mutation; Generations; Genes; Genetic; Genetic Models; Goals; Growth; Head; Human; Human Genetics; Human Pathology; Intellectual functioning disability; Kinetochores; Knockout Mice; Lead; Life; Link; Malignant Neoplasms; Mental Retardation; Mentors; Microcephaly; Mitosis; Mitotic spindle; Modeling; Mus; Mutate; Mutation; Nerve Degeneration; Neurodevelopmental Disorder; Neurons; Neurosciences; Nuclear Pore; Nuclear Pore Complex; Nucleotides; Patients; Phase; Phenocopy; Phenotype; Population; Primates; Proteins; Protocols documentation; Research; Research Personnel; Rodent Model; Role; Severities; Stem cells; Structure; Techniques; Testing; Time; Tissues; Transgenic Mice; base; career development; cellular imaging; human disease; in utero; in vitro Model; in vivo; induced pluripotent stem cell; insight; member; molecular pathology; mouse model; nerve stem cell; nervous system disorder; neurogenesis; neuroregulation; new technology; novel; nuclear pore complex protein p107; premature; progenitor; relating to nervous system; research study; self-renewal; stem cell differentiation

PROJECT SUMMARY/ABSTRACT The combination of human genetics, animal models and the recent addition of induced pluipotent stem cells (iPSCs) to the human neurodevelopmental disease modeling toolbox has the potential to greatly expand our understanding of human disease mechanisms. To date, a major challenge to understanding human neurodevelopmental disorders has been the lack of affected tissue. The capacity of iPSCs to differentiate the full complement of neural tissue, from neural progentirors (NP) to mature cortical neurons, from patient iPSC opens an exciting new avenue to understanding unique human features of disease. In this application, I propose to tailor this new technology to model defects in neurogenesis which underlie autosomal recessive primary microcephaly (MCPH). MCPH is a neurodevelopmental disorder characterized by a great reduction of head growth in utero and is accompanied by nonprogressive mental retardation. MCPH is the result of cerebral cortex hypoplasia and generalized diminution of an otherwise architecturally normal brain, a phenotype that is thought to result from defective NP proliferation early in development. MCPH is well suited for this new modeling approach as NPs differentiate early in iPSC differentiation protocols and proliferation can be evaluated within the context of neural rosettes. To correlate iPSCs in vitro modeling data to in vivo brain development, I propose to utilize a combination of patient iPSCs, transgenic mouse iPSCs and animal models. To test the sensitivity of iPSC modeling to convey unique mechanistic information, I propose to model two genetic causes for MCPH that should perturb the same set of cells in different ways or with varying severity. To gain a more comprehensive understanding of the role of in neurogenesis in the molecular pathology of MCPH according to the techniques described above I am proposing to model both Nucleoporin 107 (NUP107) and abnormal spindle-like microcephaly associated (ASPM). I recently identified Nucleoporin 107 (NUP107), a gene not previously linked to human disease, as a causative gene for MCPH. To pursue the proposed research, I have generated iPSCs from Nup107 patient and control fibroblasts and chimeric mice for a conditional NUP107 gene trap allele (NUP107GT). I have also identified a novel ASPM mutation, the gene most commonly mutated in MCPH, for which iPSC modeling will be pursued as an independent investigator. The level of mechanistic understanding that can be gained from this modeling approach for MCPH will lay the foundation that can lead to new therapies and insights into how the normal human brain develops.

项目概要/摘要 人类遗传学、动物模型和最近添加的诱导多能干细胞的结合 （iPSC）到人类神经发育疾病建模工具箱有可能极大地扩展我们的研究 了解人类疾病机制。迄今为止，理解人类的一个重大挑战 神经发育障碍一直缺乏受影响的组织。 iPSC 区分不同细胞的能力 来自患者 iPSC 的完整神经组织，从神经祖细胞 (NP) 到成熟的皮质神经元 为了解人类疾病的独特特征开辟了一条令人兴奋的新途径。在这个应用程序中，我 提议定制这项新技术来模拟常染色体隐性遗传的神经发生缺陷 原发性小头畸形（MCPH）。 MCPH 是一种神经发育障碍，其特征是 头部在子宫内生长并伴有非进行性智力低下。 MCPH 是脑部疾病的结果 皮质发育不全和其他结构正常的大脑的普遍缩小，这种表型是 被认为是由于发育早期的 NP 增殖缺陷造成的。 MCPH 非常适合这种新的 NP 在 iPSC 分化方案中早期分化和增殖的建模方法可以 在神经玫瑰花结的背景下进行评估。将 iPSC 体外建模数据与体内大脑相关联 在开发过程中，我建议结合使用患者 iPSC、转基因小鼠 iPSC 和动物模型。 为了测试 iPSC 建模传达独特机械信息的敏感性，我建议对两个模型进行建模 MCPH 的遗传原因会以不同的方式或不同的严重程度扰乱同一组细胞。 更全面地了解分子病理学中神经发生的作用 根据上述技术，我建议对核孔蛋白 107 进行建模 （NUP107）和异常纺锤状小头畸形相关（ASPM）。我最近发现了核孔蛋白 107 (NUP107)，一种以前与人类疾病无关的基因，作为 MCPH 的致病基因。为追求 根据拟议的研究，我从 Nup107 患者和对照成纤维细胞和嵌合小鼠中生成了 iPSC 条件性 NUP107 基因陷阱等位基因 (NUP107GT)。我还发现了一种新的 ASPM 突变，即基因 MCPH 中最常见的突变，将作为独立研究者对其进行 iPSC 建模。 从这种 MCPH 建模方法中获得的机械理解水平将奠定 奠定了基础，可以带来新的疗法和对正常人脑如何发育的见解。