Generation of gene-corrected oligodendrocyte progenitor cells for autologus treat

描述(申请人提供)：中枢神经系统的髓鞘形成仍然是治疗多发性硬化症和脑白质营养不良等疾病的一个难以实现的目标。最近的一些研究已经奠定了基础，并催化了目前的热情，使用谱系转换作为一种潜在的再生治疗许多神经疾病。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有明显的局限性，因此可扩增的体细胞前体细胞是基于细胞的治疗的一个追逐目标。正是随着基于细胞的疗法的出现，自体衍生的少突胶质前体细胞有望有望治疗人类脑白质营养不良。因此，临床上需要从非神经细胞中分离出自体少突胶质细胞。然而，在遗传性髓鞘变性疾病中，功能性少突胶质前体细胞不能直接从非神经细胞重新编程。这一问题揭示了基因治疗与基于细胞的治疗相结合的必要性，以治疗与遗传有关的髓鞘疾病。我假设髓鞘碱性蛋白缺陷小鼠(Shiverer，Mbpshi/SHI)，一种人类先天性脑白质营养不良的模型，可以通过移植来自成纤维细胞的自体来源的、基因修正的少突胶质前体细胞和诱导的多能干细胞(IPS)来拯救中枢神经系统的髓鞘形成。为了解决这一假设，我建议(1)从寒战(Mbpshi/SHI)成纤维细胞中产生基因校正的、功能诱导的少突胶质前体细胞，以及(2)基因校正的、直接分化为功能性少突胶质前体细胞的寒战(Mbpshi/SHI)iPS细胞。最近的进展表明，转录因子的调节成功地将患者特定的成纤维细胞重新编程为感兴趣的细胞类型。有了这些知识，我可以通过调节转录因子来改变细胞的状态，从而产生一批少突胶质前体细胞或诱导的多能干细胞，这些干细胞具有直接分化为少突胶质前体细胞的能力。此外，我建议通过引入包含完整髓鞘碱性蛋白基因及其内源调节元件的细菌人工染色体来从基因上纠正自体起始种群。通过这些方法，我建议产生一组基因校正的自体衍生的少突胶质前体细胞，当在体内注射到宿主颤抖(Mbpshi/SHI)小鼠模型中时，导致一度髓鞘减少的中枢神经系统的功能性髓鞘化。因此，首次可以产生患者特有的少突胶质前体细胞群体，这些细胞被设计为缺乏致病突变，用于潜在的遗传相关髓鞘疾病的治疗。

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.