Generation of gene-corrected oligodendrocyte progenitor cells for autologus treat

描述（由申请人提供）：中枢神经系统的髓鞘形成仍然是治疗诸如多发性硬化症和脑白质营养不良的疾病的难以实现的目标。最近的一些研究奠定了基础，并催化目前的热情使用谱系转换作为一个潜在的再生治疗许多神经系统疾病。然而，完全成熟的细胞如神经元和少突胶质细胞的移植和功能整合具有明显的局限性，因此可扩增的体细胞祖细胞是基于细胞的疗法的目标。随着基于细胞的疗法的出现，自体来源的少突胶质细胞祖细胞有望用于潜在地治疗人类脑白质营养不良。由于这个原因，存在从非神经细胞衍生自体少突胶质细胞的临床需要。然而，在遗传相关的髓鞘形成障碍疾病中，功能性少突胶质细胞祖细胞不能直接从非神经细胞重编程。这一问题揭示了基因治疗与基于细胞的治疗相结合用于治疗遗传相关的髓鞘疾病的需要。我假设，髓鞘碱性蛋白缺陷小鼠（shiverer，Mbpshi/shi），人类先天性脑白质营养不良的模型，可以通过移植自体来源的，基因校正的少突胶质细胞祖细胞从成纤维细胞和诱导多能干细胞（iPS）的中枢神经系统髓鞘形成的拯救。为了解决这一假设，我提出（1）从shiverer（Mbpshi/shi）成纤维细胞产生基因校正的、功能性诱导的少突胶质细胞祖细胞，和（2）基因校正的和直接分化shiverer（Mbpshi/shi）iPS细胞为功能性少突胶质细胞祖细胞。最近的进展已经证明成功地调节转录因子谱以将患者特异性成纤维细胞重编程为感兴趣的细胞类型。有了这些知识，我可以调节转录因子谱，改变细胞的状态，产生一群少突胶质细胞祖细胞或诱导多能干细胞，它们具有直接分化为少突胶质细胞祖细胞的能力。此外，我建议通过引入一个细菌人工染色体，包含完整的髓鞘碱性蛋白基因，作为其内源性调控元件的威尔斯遗传校正自体起始人群。通过这些方法，我建议产生一个人口的基因校正autologously衍生的少突胶质细胞祖细胞，当在体内注射到主机shiverer（Mbpshi/shi）小鼠模型导致功能髓鞘的一次hypomyelinated中枢神经系统。因此，第一次可以产生患者特异性的少突胶质细胞祖细胞群，其已经被工程化以缺乏致病突变，用于遗传相关的髓鞘疾病的潜在治疗。

DESCRIPTION (provided by applicant): Myelination of the central nervous system remains an elusive goal for treating disorders such as multiple sclerosis and leukodystrophies. A number of recent studies have laid the foundation and catalyzed current enthusiasm for the use of lineage conversion as a potential regenerative therapy for many neurological disorders. However, transplantation and functional integration of fully mature cells such as neurons and oligodendrocytes has clear limitations, therefore expandable somatic progenitors are a sought after target for cell-based therapies. It is with the advent of cell based therapies that autologously derived oligodendrocyte progenitor cells hold promise for potentially treating human leukodystrophies. For this reason, there exists a clinical need to derive autologous oligodendrocytes from non-neural cells. However, in genetic related dysmyelinating disease, functional oligodendrocyte progenitor cells cannot be directly reprogrammed from non- neural cells. This issue brings to light the need for gene-therapy coupled with cell based therapies for treating genetic related myelin diseases. I hypothesized that myelin basic protein deficient mice (shiverer, Mbpshi/shi), a model of human congenital leukodystrophies, can be rescued through transplantation of autologously derived, gene corrected oligodendrocyte progenitor cells from fibroblasts and induced pluripotent stem (iPS) cells for the myelination of the central nervous system. To address this hypothesis I propose to (1) generate gene- corrected, functional induced oligodendrocyte progenitor cells from shiverer (Mbpshi/shi) fibroblasts and (2) gene-correct and directly differentiate shiverer (Mbpshi/shi) iPS cells to functional oligodendrocyte progenitor cells. Recent advances have demonstrated the successful modulation of transcription factor profiles to reprogram patient specific fibroblasts into cell types of interest. With this knowledge I can modulate transcription factor profiles to change the state of the cell to generate a population of oligodendrocyte progenitor cells or induced pluripotent stem cells which have the capability of being directly differentiated into oligodendrocyte progenitor cells. In addition, I propose to genetically correct the autologous starting populations by introducing a bacterial artificial chromosome that contains the full myelin basic protein gene, as wells as its endogenous regulatory elements. Through these methods I propose to generate a population of gene-corrected autologously derived oligodendrocyte progenitor cells that when injected in vivo into the host shiverer (Mbpshi/shi) mouse model leads to the functional myelination of the once hypomyelinated central nervous system. Thus for the first time a patient-specific population of oligodendrocyte progenitor cells can be generated, that have been engineered to lack disease causing mutations, for the potential treatment of genetic related myelin diseases.