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化仍然是治疗多发性硬化症和白细胞营养不良症等疾病的难以捉摸的目标。最近的许多研究奠定了基础，并促进了目前对使用谱系转化为许多神经系统疾病的潜在再生疗法的热情。但是，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，因此可扩展的体祖细胞是基于细胞的疗法的靶标。自体衍生的少突胶质细胞祖细胞的出现，基于细胞的疗法有望有望治疗人类白细胞营养不良。因此，存在从非神经细胞中得出自体少突胶质细胞的临床需求。然而，在遗传相关的肿瘤疾病中，功能性少突胶质细胞祖细胞不能直接从非神经细胞中重新编程。这个问题揭示了对基因疗法的需求，以及用于治疗遗传相关髓鞘疾病的基于细胞的疗法。 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.为了解决这一假设，我提出（1）生成了来自发抖者（MBPSHI/SHI）成纤维细胞的基因校正的，功能诱导的少突胶质细胞祖细胞，以及（2）基因 - 校正，并直接区分湿度器（MBPSHI/SHI）IPS（MBPSHI/SHI）IPS细胞（MBPSHI/SHI）对功能型级别的元素细胞。最近的进步表明，转录因子曲线的成功调节，以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曲线以改变细胞的状态，以产生少突胶质细胞祖细胞的种群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细胞祖细胞的能力。此外，我建议通过引入包含完整的髓磷脂碱性蛋白基因的细菌人工染色体以及其内源性调节元素来纠正自体开始种群。通过这些方法，我建议生成一组基因校正的自体衍生的少突胶质细胞祖细胞，这些细胞将体内注射到宿主颤抖者（MBPSHI/SHI）小鼠模型中会导致曾经降低了肾上腺素降低的中枢神经系统的功能性髓鞘化。因此，这是第一次可以产生特定于患者的少突胶质细胞祖细胞，这些细胞已被设计为缺乏疾病引起突变，以治疗遗传相关的髓磷脂疾病。