Therapeutic Potential of Embryonic Stem Cell-Derived Myogenic Precursors

胚胎干细胞衍生的肌源性前体的治疗潜力

• 批准号: 7799717
• 负责人: BAOLI YANG
• 金额: $ 30.69万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7799717
• 关键词: Acute; Adult; Affect; Animal Model; Aorta; Bioluminescence; Blood Vessels; Cell Differentiation process; Cell Proliferation; Cell Survival; Cell Transplantation; Cell Transplants; Cell surface; Cells; Characteristics; Clinical; Coculture Techniques; Collaborations; Development; Disease; Dorsal; Embryo; Engraftment; Feline Immunodeficiency Virus; Fetal Tissues; Future; Gene Expression Profile; Generations; Genes; Goals; Histologic; Homing; Human; Immunocompetent; In Vitro; Individual; Inherited; Intra-Arterial Injections; Knockout Mice; Label; Laboratories; Limb-Girdle Muscular Dystrophies; Long-Term Effects; Luciferases; Mediating; Methods; Modeling; Molecular; Molecular Probes; Monitor; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscle Weakness; Muscular Dystrophies; Myoblasts; Myopathy; Natural regeneration; Nature; Normal Cell; Pattern; Phenotype; Population; Positioning Attribute; Property; Protocols documentation; Research; Rest; Sarcoglycans; Severities; Site; Skeletal Muscle; Source; Stem cell transplant; Stem cells; Surface; Surface Antigens; System; Testing; Therapeutic; Therapeutic Agents; Transplantation; base; cell type; embryonic stem cell; functional improvement; gene therapy; human embryonic stem cell; human embryonic stem cell line; in vivo; internal control; mouse model; muscle regeneration; precursor cell; progenitor; promoter; reconstitution; regenerative; repaired; stem cell differentiation; therapeutic gene; trafficking; vector; wasting

Muscular dystrophies (MDs) are a broad group of inherited diseases that are characterized by progressive skeletal muscle weakness and wasting with a variable clinical course. The molecular mechanisms whereby individual skeletal muscle fibers undergo degeneration have not been well defined, and there are still no cures for these diseases. This proposal explores the therapeutic potential of mouse embryonic stem (ES) cells, with the ultimate goal of discovering principles that govern, the differentiation of ES cells into myogenic precursor cells that can be used in cell transplantation in murine models of MDs. To date, there has been no definitive demonstration that stem cells that arise in vitro during differentiation into embryoid bodies (EBs) are capable of engrafting either normal or dystrophic adult muscle. In collaboration with Cossu and colleagues, the Campbell laboratory showed that mesoangioblasts, stem cells isolated from embryonic dorsal aorta, can correct the phenotype in mouse model of MD. We are thus well positioned to investigate the nature of the common vascular-myogenic progenitor that can be formed in EBs, and the mechanisms governing the potential for such cells to engraft in adult myogenic microenvironments. We are proposing methods for isolating enriched populations of EB-derived progenitors using either selectable markers driven by myogenic specific promoters or cell purification via surface markers, and will endeavor to more carefully define the surface antigen phenotype and in vivo properties of this population of cells. We will test the hypotheses that engraftment is facilitated by homing, enhanced cell survival, or cell proliferation, and probe the molecular basis for these observations using a non-invasive bioluminescence monitoring system that will also allow us to investigate the long-term effect of cell-mediated therapy. Our results will enable methods for enhanced myogenic development from ES cells and reconstitution of the adult myogenic system as a model for myogenic research and cellular therapies. Future efforts will explore the similarities and differences in the myogenic potential of murine and human ES cells, and the therapeutic potential of in vitro differentiated human ES cells.

肌营养不良症 (MD) 是一类广泛的遗传性疾病，其特征是进行性骨骼肌无力和萎缩，临床病程各异。个体骨骼肌纤维发生变性的分子机制尚未明确，并且仍然无法治愈这些疾病。该提案探讨了小鼠胚胎干 (ES) 细胞的治疗潜力，最终目标是发现 ES 细胞分化为肌源性前体细胞的原理，这些细胞可用于 MD 小鼠模型的细胞移植。迄今为止，还没有明确的证据表明干细胞在分化为胚状体（EB）的过程中在体外产生 能够移植正常或营养不良的成人肌肉。坎贝尔实验室与 Cossu 及其同事合作，表明中成血管细胞（从胚胎背主动脉分离的干细胞）可以纠正 MD 小鼠模型的表型。因此，我们有能力研究 EB 中形成的常见血管肌源祖细胞的性质，以及控制此类细胞在成体肌源性微环境中移植的潜力的机制。我们正在提出使用由生肌特异性启动子驱动的选择性标记或通过表面标记进行细胞纯化来分离丰富的 EB 衍生祖细胞群的方法，并将努力更仔细地定义该细胞群的表面抗原表型和体内特性。我们将测试归巢、增强细胞存活或细胞增殖促进植入的假设，并探究 使用非侵入性生物发光监测系统进行这些观察的分子基础也将使我们能够研究细胞介导疗法的长期效果。我们的研究结果将为增强 ES 细胞的肌源性发育和重建成体肌源性系统作为肌源性研究和细胞治疗的模型提供方法。未来的工作将探索小鼠和人类 ES 细胞的生肌潜力的异同，以及体外分化的人类 ES 细胞的治疗潜力。