Delivery of human muscle SP cells to dystrophic muscle

将人类肌肉 SP 细胞递送至营养不良的肌肉

• 批准号: 6824663
• 负责人: EMANUELA GUSSONI
• 金额: $ 26.93万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6824663
• 关键词: CD antigens; CD34 molecule; NOD mouse; SCID mouse; cell differentiation; cell population study; cell sorting; cell transplantation; clinical research; flow cytometry; fluorescent in situ hybridization; genetically modified animals; hematopoiesis; human fetus tissue; human tissue; immunocytochemistry; intraarterial administration; intramuscular injections; intravenous administration; muscle cells; muscular dystrophy; myogenesis; postmortem; therapy design /development; tissue /cell preparation

DESCRIPTION (provided by applicant): Muscle SP cells are a heterogeneous population of primitive cells that demonstrated hematopoietic and myogenic differentiation potential in vivo. First discovered in mice, these cells hold promise for optimization of cell-based therapy for muscular dystrophy. To date, little is known about human-derived muscle SP cells and their presumed clinical potential. The goals of this proposal are to study the differentiation potential of human fetal and adult-derived muscle SP cells, and to optimize their use in pre-clinical experiments using newly generated muscular dystrophy mouse models. These goals will be achieved via the following specific aims: Aim 1.1. Study the potential of unfractionated human muscle-derived SP and MP cells to differentiate into myogenic or hematopoietic cells in vitro. Human samples will be obtained from autopsy fetal and adult tissues and from adult-derived discarded surgical tissue. Aim 1.2. Fractionate human fetal and adult muscle SP and MP cells based on the expression of the cell surface antigens CD34, CD90 and CD133. Compare the ability of fractionated versus unfractionated cells to differentiate into myogenic or hematopoietic cells in vitro. Aim 2.1. Compare the ability of unfractionated and fractionated muscle-derived SP and MP cells to fuse into dystrophic myofibers in vivo. Perform intramuscular injections into NOD/RAG1/null-DMD/mdx5cv mice using the fetal and adult-derived human muscle SP and MP cell fractions enriched for myogenic precursors (Aim 1.2). Aim 2.2. Assess the clinical potential of human fetal and adult-derived muscle SP or MP cells with myogenic activity to engraft dystrophic skeletal muscles in vivo. Perform intra-arterial and intravenous injections of human muscle SP or MP cells in non-irradiated NOD/RAG1/null-DMD/mdx5cv. Aim 2.3. Assess the ability of fractionated human fetal and adult-derived muscle SP or MP cells to differentiate into hematopoietic cells in vivo. Inject unfractionated and fractionated cells with hematopoietic activity (Aims 1.1 and 1.2) into the circulation of lethally irradiated NOD/RAG1/null/mdx5cv or NOD/RAG1/null-Pfp/null DMD/mdx5cv mice. These studies will enhance our basic knowledge on human muscle SP cells, their differentiation potential in fetal and adult tissue, and will evaluate their therapeutic promise for muscular dystrophy.

描述（由申请人提供）：肌肉 SP 细胞是原始细胞的异质群体，在体内表现出造血和肌原性分化潜力。这些细胞首次在小鼠中发现，有望优化肌营养不良症的细胞疗法。迄今为止，人们对人源性肌肉 SP 细胞及其推测的临床潜力知之甚少。该提案的目标是研究人类胎儿和成人来源的肌肉 SP 细胞的分化潜力，并使用新生成的肌营养不良小鼠模型优化其在临床前实验中的使用。这些目标将通过以下具体目标来实现： 目标 1.1。研究未分级的人肌肉来源的 SP 和 MP 细胞在体外分化为肌原细胞或造血细胞的潜力。人类样本将从尸检胎儿和成人组织以及成人丢弃的手术组织中获得。 目标 1.2。根据细胞表面抗原 CD34、CD90 和 CD133 的表达对人类胎儿和成人肌肉 SP 和 MP 细胞进行分级。比较分级细胞与未分级细胞在体外分化为肌原细胞或造血细胞的能力。 目标 2.1。比较未分级和分级的肌肉来源的 SP 和 MP 细胞在体内融合成营养不良的肌纤维的能力。使用富含肌源性前体的胎儿和成人来源的人类肌肉 SP 和 MP 细胞分数对 NOD/RAG1/null-DMD/mdx5cv 小鼠进行肌内注射（目标 1.2）。 目标 2.2。评估具有生肌活性的人类胎儿和成人来源的肌肉 SP 或 MP 细胞在体内移植营养不良的骨骼肌的临床潜力。在未照射的 NOD/RAG1/null-DMD/mdx5cv 中进行人体肌肉 SP 或 MP 细胞的动脉内和静脉内注射。 目标 2.3。评估分馏的人类胎儿和成人来源的肌肉 SP 或 MP 细胞在体内分化为造血细胞的能力。将具有造血活性的未分级和分级细胞（目标 1.1 和 1.2）注入致死照射的 NOD/RAG1/null/mdx5cv 或 NOD/RAG1/null-Pfp/null DMD/mdx5cv 小鼠的循环中。 这些研究将增强我们对人类肌肉 SP 细胞及其在胎儿和成人组织中的分化潜力的基础知识，并将评估它们对肌营养不良症的治疗前景。