Human iPSC and glial chimeric modeling of Pelizaeus-Merzbacher Disease

 描述(申请人提供)：Pelizaeus-Merzbacher病(PMD)是一种严重的X连锁儿童神经退行性疾病，影响中枢神经系统的髓鞘形成。PMD是由PLP1基因突变引起的，PLP1基因编码中枢神经系统最常见的髓鞘蛋白。PMD表现出一系列临床表型，反映了广泛的基因异质性，但几乎所有形式的疾病都会导致进行性神经恶化和死亡，通常发生在儿童时期。随着诱导多能干细胞(IPSC)技术和细胞命运重组的出现，我们现在已经开发出了强大的方法来在体外产生患者特异性的少突胶质细胞。这提供了新的途径来模拟PMD内的表型和基因型谱。我们已经从PMD患者那里生成了一个大型的综合性IPSC品系小组，涵盖了该疾病的全部基因型谱和表型谱。在这项应用中，我们将重点研究PMD患者少突胶质细胞中特定于患者的分子和细胞缺陷，并为治疗提供测试平台。目的1将使用我们开发的强大的人类IPSC来源的少突胶质细胞分化方案，作为检测PMD患者少突胶质细胞功能障碍的表型平台，并评估临床相关药物治疗在体外拯救个体表型的能力。目的2将使用一种方法，通过这种方法，我们可以制造出人源化的嵌合小鼠，其少突胶质细胞和髓鞘在很大程度上已被人的iPSC来源的少突胶质细胞取代。将这项技术与我们面板中的PMD iPSC系相结合，将使我们能够产生针对患者的PMD神经胶质嵌合体，并在体内检测少突胶质细胞功能障碍。在目标3中，我们将测试转基因PMD IPSC来源的OPC是否在神经胶质嵌合小鼠中显示出恢复的髓鞘形成能力。总之，这些研究应该为我们提供对推动PMD发病机制的细胞和分子缺陷的新见解。我们的目标是为PMD的建模和治疗建立体外和体内平台，同时为髓鞘遗传性疾病的治疗提供广泛的策略。

 DESCRIPTION (provided by applicant): Pelizaeus-Merzbacher disease (PMD) is a severe X-linked pediatric neurodegenerative disorder impacting myelination in the central nervous system. PMD results from mutations in the PLP1 gene which encodes the most prevalent myelin protein of the central nervous system. PMD exhibits a spectrum of clinical phenotypes that reflect wide genotypic heterogeneity, but nearly all forms of the disease result in progressive neurological deterioration and death, often during childhood. With the advent of induced pluripotent stem cell (iPSC) technologies and cell fate reengineering, we have now developed robust methods for generation of patient- specific oligodendrocytes in vitro. This provides new access to model the phenotypic and genotypic spectra within PMD. We have generated a large comprehensive panel of characterized iPSC lines from PMD patients that spans the full genotypic and phenotypic spectrum of the disorder. In this application we will focus on the patient-specific molecular and cellular deficits in PMD patient oligodendrocytes and provide platforms for testing of therapeutics. Aim 1 will use the robust human iPSC-derived oligodendrocyte differentiation protocols that we have developed as a phenotyping platform for examining PMD patient oligodendrocyte dysfunction and assessing the ability of clinically-relevant pharmacologic therapies to rescue individual phenotypes in vitro. Aim 2 will use an approach by which we can produce humanized chimeric mice whose oligodendrocytes and myelin have been largely replaced by human iPSC-derived oligodendroglia. Combining this technology with PMD iPSC lines from our panel will enable us to generate patient-specific PMD glial chimeras and examine oligodendrocyte dysfunction in vivo. In Aim 3 we will test if genetically-corrected PMD iPSC-derived OPCs show restored myelination capacity in glial chimeric mice. Together, these studies should provide us new insight into the cell and molecular deficits driving the pathogenesis of PMD. Our goal is to establish in vitro and in vivo platforms for the modeling and treatment of PMD, while providing a broad strategy for the treatment of hereditary disorders of myelin.