Generation of gene-corrected oligodendrocyte progenitor cells for autologus treat

用于自体治疗的基因校正少突胶质细胞祖细胞的产生

• 批准号: 8524663
• 负责人: Angela Lager
• 金额: $ 4.22万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8524663
• 关键词: Achievement; Address; Affect; Animal Model; Autoimmune Diseases; Autologous; Autologous Transplantation; Axon; Bacterial Artificial Chromosomes; Biotechnology; Cardiac Myocytes; Cell Therapy; Cells; Clinical; Clinical Treatment; Conditioned Culture Media; Coupled; Data; Differentiated Gene; Disease; Engineering; Fibroblasts; Foundations; Generations; Genes; Genetic; Goals; Hepatocyte; Hereditary Disease; Human; In Vitro; Individual; Knowledge; Length; Methods; Modeling; Multiple Sclerosis; Mus; Myelin; Myelin Basic Proteins; Nature; Neuraxis; Neurons; Oligodendroglia; Patients; Pluripotent Stem Cells; Population; Property; Publishing; Regulatory Element; Scientific Advances and Accomplishments; Shivering; Skin; Source; Spastic Paraplegia; Stem cells; System; Therapeutic; Time; Transplantation; Work; cell type; disease-causing mutation; dysmyelination; functional improvement; gene correction; gene therapy; in vivo; induced pluripotent stem cell; interest; leukodystrophy; mouse model; myelination; nervous system disorder; oligodendrocyte lineage; progenitor; public health relevance; regenerative therapy; standard care; transcription factor

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.

描述（由申请人提供）：中枢神经系统的髓鞘形成对于治疗多发性硬化症和脑白质营养不良等疾病仍然是一个难以捉摸的目标。最近的许多研究为使用谱系转换作为许多神经系统疾病的潜在再生疗法奠定了基础并激发了当前的热情。然而，完全成熟的细胞（例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基于细胞的疗法的追捧目标。正是随着基于细胞的疗法的出现，自体衍生的少突胶质细胞祖细胞有望用于治疗人类脑白质营养不良。因此，临床上存在从非神经细胞衍生自体少突胶质细胞的需要。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由非神经细胞重编程。这个问题揭示了基因疗法与细胞疗法相结合来治疗遗传相关髓磷脂疾病的需求。我假设髓磷脂碱性蛋白缺陷小鼠（shiverer，Mbpshi/shi）是一种人类先天性脑白质营养不良模型，可以通过移植来自成纤维细胞的自体来源的、基因校正的少突胶质细胞祖细胞和用于髓鞘形成的诱导多能干（iPS）细胞来挽救中枢神经系统的。为了解决这一假设，我建议（1）从颤抖者（Mbpshi/shi）成纤维细胞中产生基因校正的功能性诱导少突胶质细胞祖细胞，以及（2）基因校正并直接将颤抖者（Mbpshi/shi）iPS细胞分化为功能性少突胶质细胞祖细胞细胞。最近的进展表明，可以成功地调节转录因子谱，将患者特异性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或具有直接分化为少突胶质细胞祖细胞能力的诱导多能干细胞。此外，我建议通过引入包含完整髓磷脂碱性蛋白基因及其内源性调控元件的细菌人工染色体来对自体起始群体进行遗传纠正。通过这些方法，我建议产生一群基因校正的自体衍生的少突胶质细胞祖细胞，当将其注射到宿主颤抖者（Mbpshi/shi）小鼠模型体内时，会导致曾经髓鞘发育不良的中枢神经系统功能性髓鞘形成。因此，首次可以产生患者特异性的少突胶质细胞祖细胞群，这些祖细胞经过改造，缺乏引起疾病的突变，可用于遗传相关髓磷脂疾病的潜在治疗。