Generation of gene-corrected oligodendrocyte progenitor cells for autologus treat

描述（由申请人提供）：中枢神经系统髓鞘形成仍然是治疗多发性硬化症和白质营养不良等疾病的难以捉摸的目标。最近的一些研究已经奠定了基础，并催化了目前使用谱系转换作为许多神经系统疾病的潜在再生疗法的热情。然而，完全成熟细胞（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，因此可扩展的体细胞祖细胞是细胞治疗的追求目标。随着细胞疗法的出现，自体少突胶质细胞祖细胞有望治疗人类白质营养不良。因此，临床需要从非神经细胞中提取自体少突胶质细胞。然而，在遗传相关的髓鞘异常疾病中，功能性少突胶质细胞祖细胞不能从非神经细胞直接重编程。这一问题揭示了基因疗法结合细胞疗法治疗遗传相关髓磷脂疾病的必要性。我假设髓鞘碱性蛋白缺陷小鼠（shiverer, Mbpshi/shi），一种人类先天性白质营养不良的模型，可以通过移植自源的、基因校正的少突胶质细胞祖细胞和用于中枢神经系统髓鞘形成的诱导多能干细胞（iPS）来拯救。为了解决这一假设，我建议：(1)从shiverer （Mbpshi/shi）成纤维细胞中产生基因校正的，功能诱导的少突胶质细胞祖细胞；(2)基因校正并直接将shiverer (Mbpshi/shi) iPS细胞分化为功能少突胶质细胞祖细胞。最近的进展已经证明了转录因子谱的成功调节，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来改变细胞的状态，从而产生少突胶质细胞祖细胞群体或诱导多能干细胞，这些干细胞具有直接分化为少突胶质细胞祖细胞的能力。此外，我建议通过引入含有完整髓鞘碱性蛋白基因及其内源性调控元件的细菌人工染色体，对自体起始种群进行遗传校正。通过这些方法，我建议产生一群基因校正的自体来源的少突胶质细胞祖细胞，当注射到宿主shiverer （Mbpshi/shi）小鼠模型体内时，导致曾经的低髓鞘中枢神经系统的功能髓鞘形成。因此，首次可以产生患者特异性的少突胶质细胞祖细胞群，这些细胞已被改造成缺乏引起疾病的突变，用于遗传相关髓磷脂疾病的潜在治疗。

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.