Systemic delivery of muscle stem cell for muscle disease therapy

肌肉干细胞的全身递送用于肌肉疾病治疗

基本信息

批准号：
10451411
负责人：
ATSUSHI ASAKURA
金额：
$ 20.46万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10451411
关键词：
Achievement Affect Animal Model Autologous B-Lymphocytes Blood Circulation Blood Vessels Capillary Endothelial Cell Cell Nucleus Cell Survival Cell Therapy Cell Transplantation Cells Chronic Clinical Trials Coculture Techniques Code Communication Disease Duchenne muscular dystrophy Dystrophin Encapsulated Engraftment Extravasation Failure Gene Proteins Genes Genetic Goals Human Immune Injections Intra-Arterial Injections Intramuscular Injections Lead Lipids Mannitol Mediating Mediator of activation protein Membrane Methods Microscope Modification Mus Muscle Muscle Contraction Muscle Fibers Muscle satellite cell Muscular Dystrophies Mutation Myoblasts Myopathy Natural regeneration Nucleic Acids Optics Outcome Patients Proliferating Proteins Protocols documentation Quality of life Research Resolution Sarcolemma Scheme Skeletal Muscle Skeletal Muscle Satellite Cells Source Stem cell transplant Structure System Testing Therapeutic Thinness Three-Dimensional Imaging Tissues Transduction Gene Transfection Transgenes Transplantation Vascular Permeabilities cell type cremaster muscle extracellular vesicles improved induced pluripotent stem cell injured insight mdx mouse micro-dystrophin migration muscle degeneration muscle regeneration muscular dystrophy mouse model nanoparticle novel therapeutic intervention overexpression practical application precursor cell prevent programs real-time images receptor repaired restoration satellite cell scale up self-renewal stem cells vector

项目摘要

Abstract Duchenne Muscular Dystrophy (DMD) is caused by mutations in the gene coding for dystrophin, which functions to maintain muscle fiber structure and function in the whole body by preventing it from being damaged during muscle contraction. Presently, there is no definitive treatment for DMD patients, and current therapies focus on prolonging survival and improving quality of life. Definitive treatment will require that functional dystrophin protein is restored in all affected muscle groups. Possible approaches to restoring dystrophin expression in muscle fibers include cell therapy.We hope that transplantation of a particular type of cell capable of regenerating muscle will help us to develop new therapeutic approaches to this disease. Muscle stem cells, termed satellite cells, isolated from healthy donors or patients should be able to provide dystrophin and repair muscle damage in DMD patients. For efficient DMD therapy, satellite cells which maintain the ability to self-renew are also necessary. However, satellite cells are rare (2-7% of all muscle nuclei) and often difficult to isolate. In addition, efficient myogenic differentiation and the scale-up of myogenic differentiation remain elusive and must be developed further in order to generate effective cell-based therapy for chronic muscle diseases. Patient-derived induced pluripotent stem cells (iPSCs) are the ideal cell source to obtain an unlimited number of myogenic cells that escape immune rejection after engraftment. However, the failure of systemic delivery of the injected cells has hindered practical application in patients. Our long-term goals are to develop an effective supply of satellite cells for cell therapy, to find an efficient systemic delivery method that can treat all affected muscles, and to enable the injected cells to self-renew and reach an effective mass. The specific aims of this application: 1. To determine possible interactions between CD24-expressing satellite cells and capillary endothelial cells via real-time imaging and 3- D imaging after our recently established whole muscle tissue clearing protocol. 2. To develop systemic delivery methods for satellite cells and hiPSC-derived myogenic progenitor cells in combination with genetic modification, including transduction of an extravasation factor, CD24, and Mannitol, an agent for enhancing vascular permeabilization. 3. To develop systemic delivery methods for satellite cells via Extracellular Vesicle (EV)- mediated CD24 transfer. In combination with gene and protein transduction, this concerted approach will help us to make satellite cells that can be systemically transplanted into patients for a definitive cure of DMD. The proposed specific aims will test whether systemically-injected satellite cells can be delivered to a target injured muscle and contribute to muscle regeneration as well as to the satellite cell pool. The anticipated outcome of the proposed specific aims will provide valuable insights for satellite cell-based cell therapies in muscular dystrophies and other muscle diseases. Eventually, our experimental scheme will be directly applied to human satellite cells isolated from DMD patients. The DMD-specific satellite cells stably transfected with CD24 and microdystrophin will be able to produce enough satellite cells for autologous and systemic cell transplantation to DMD patients.

抽象的 Duchenne肌肉营养不良（DMD）是由编码肌营养不良蛋白的基因中的突变引起的，该突变功能通过防止在肌肉纤维结构和功能中保持肌肉纤维结构和功能。肌肉收缩。目前，没有针对DMD患者的确切治疗方法，目前的疗法专注于延长生存和改善生活质量。确定的治疗将需要功能性肌营养不良蛋白在所有受影响的肌肉群中恢复。恢复肌肉纤维中肌营养不良蛋白表达的可能方法包括细胞疗法。我们希望移植能够再生肌肉的特定类型的细胞帮助我们开发新的治疗方法来解决这种疾病。肌肉干细胞，称为卫星细胞，分离来自健康的捐助者或患者应该能够在DMD患者中提供肌营养不良蛋白和修复肌肉损伤。对于有效的DMD疗法，还需要保持自我更新能力的卫星细胞。然而，卫星细胞很少见（占所有肌肉核的2-7％），通常很难分离。另外，有效的肌原性分化和肌源分化的扩大仍然难以捉摸，必须进一步发展为慢性肌肉疾病产生有效的基于细胞的治疗。患者衍生的诱导多能茎细胞（IPSC）是获得无限数量逃脱免疫细胞的理想细胞来源植入后拒绝。但是，注射细胞的全身输送失败已阻碍了实际的在患者中应用。我们的长期目标是开发有效的卫星细胞进行细胞治疗，以便找到一种有效的全身递送方法，可以治疗所有受影响的肌肉，并使注射细胞能够到达自我更新并达到有效的质量。此应用程序的具体目的：1。确定可能通过实时成像和3- D成像是在我们最近确定的整个肌肉组织清除方案之后。 2。开发系统交付卫星细胞和HIPSC衍生的肌源性祖细胞结合遗传修饰的方法，包括转导外渗出因子，CD24和甘露醇（增强血管的药物）透化。 3。通过细胞外囊泡（EV） - 开发卫星细胞的全身递送方法 - 介导的CD24转移。结合基因和蛋白质转导，这种协同的方法将有助于我们制作可以系统地移植到患者的DMD治疗的患者中的卫星细胞。这提出的特定目标将测试是否可以将系统注射的卫星细胞传递到目标受伤的目标肌肉并有助于肌肉再生以及卫星细胞库。预期的结果拟议的特定目标将为肌肉营养不良的卫星细胞细胞疗法提供宝贵的见解和其他肌肉疾病。最终，我们的实验方案将直接应用于人卫星细胞从DMD患者中分离出来。 DMD特异性的卫星细胞稳定地用CD24和微肺炎转染将能够产生足够的卫星细胞，以自体和全身细胞向DMD患者移植。