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

项目总结/摘要 结合人类遗传学、动物模型和最近加入的诱导多能干细胞， （iPSCs）的人类神经发育疾病建模工具箱有可能大大扩展我们的研究。 了解人类疾病的机制。迄今为止，理解人类的一个主要挑战是， 神经发育障碍一直缺乏受影响的组织。iPSCs分化成 来自患者iPSC的从神经祖细胞（NP）到成熟皮质神经元的完整神经组织 开启了一条令人兴奋的新途径，以了解人类独特的疾病特征。在本申请中，我 我建议调整这项新技术，以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，其特征在于 在子宫内头部生长，并伴有非进行性智力迟钝。MCPH是大脑 皮质发育不全和其他结构正常的大脑的普遍缩小， 被认为是由发育早期NP增殖缺陷引起的。MCPH非常适合这种新的 在iPSC分化方案和增殖中，作为NPs早期分化的建模方法可以是 在神经系统的背景下进行评估。将iPSC体外建模数据与体内脑 为了进一步开发，我建议利用患者iPSC、转基因小鼠iPSC和动物模型的组合。 为了测试iPSC建模对传达独特机制信息的敏感性，我建议建模两个 MCPH的遗传原因应该以不同的方式或以不同的严重程度干扰同一组细胞。 从分子病理学的角度更全面地了解在神经发生中的作用 根据上述技术，我建议将核孔蛋白107 （NUP 107）和异常梭形小头畸形相关（ASPM）。我最近发现核孔蛋白107 （NUP 107），一种以前与人类疾病无关的基因，作为MCPH的致病基因。追求 我已经从Nup 107患者和对照成纤维细胞以及嵌合小鼠中产生了iPSC， 条件NUP 107基因陷阱等位基因（NUP 107 GT）。我还发现了一种新的ASPM突变基因， 在MCPH中最常见突变，将作为独立研究者对其进行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奠定 该基金会可以导致新的疗法和对正常人类大脑如何发育的见解。

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.