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

项目摘要/摘要 人类遗传学、动物模型和最近增加的诱导性多能干细胞的结合 (IPSCs)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 对人类疾病机制的理解。到目前为止，理解人类的一个主要挑战 神经发育障碍一直是缺乏受影响的组织。IPSC区分不同种族的能力 完整的神经组织，从神经孕激素(NP)到成熟的皮质神经元，来自患者IPSC 为了解疾病的独特人类特征开辟了一条令人兴奋的新途径。在此应用程序中，我 建议定制这项新技术来模拟常染色体隐性遗传背后的神经发生缺陷 原发性小头畸形(MCPH)。MCPH是一种神经发育障碍，其特征是 头部在子宫内发育，并伴有非进行性精神发育迟缓。MCPH是脑出血的结果 大脑皮质发育不良和广泛性萎缩，在其他方面结构正常的大脑，这种表型是 认为是发育早期有缺陷的NP增殖所致。MCPH非常适合这一新的 当核子在IPSC分化协议和增殖的早期分化时建模的方法可以 在神经花环的背景下进行评估。IPSCs体外建模数据与体内脑的相关性 为了开发，我建议结合使用患者ipscs、转基因小鼠ipscs和动物模型。 为了测试IPSC建模传达独特机械信息的敏感度，我建议建立两个模型 MCPH的遗传原因应该以不同的方式或不同的严重程度扰乱同一组细胞。 为了更全面地了解神经发生在分子病理学中的作用 根据上述技术，我建议对核孔素107和核孔素107 (NUP107)和异常纺锤体样小头畸形(ASPM)。我最近发现了核孔素107 (NUP107)，一个以前与人类疾病无关的基因，作为MCPH的致病基因。为了追求 拟议的研究，我已经从Nup107患者和对照成纤维细胞和嵌合体小鼠中培养出IPSCs用于 一个条件性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 在MCPH中最常见的突变，将作为独立调查者进行IPSC建模。 从MCPH的这种建模方法中可以获得的机械理解水平将为 基础，可以导致新的治疗方法和对正常人脑如何发育的洞察。

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.