Engineering iPSC-Derived Skeletal Muscle and Cells for Transplantation

工程化 iPSC 衍生的骨骼肌和细胞用于移植

• 批准号: 9164844
• 负责人: ATSUSHI ASAKURA
• 金额: $ 20.14万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9164844
• 关键词: Accidents; Aging; Animal Model; Animals; Autologous; Cell Nucleus; Cell Transplantation; Cell Transplants; Cells; Cessation of life; Characteristics; Chimera organism; Chronic; Chronic Disease; Complement; Defect; Disease; Duchenne muscular dystrophy; Embryo; Embryonic Development; Engineering; Engraftment; Ethical Issues; Fetus; First Pregnancy Trimester; Generations; Goals; Human; Human Characteristics; Human body; Immune; Injection of therapeutic agent; Injury; Intraperitoneal Injections; Limb structure; Locomotion; Metabolism; Methods; Molecular Profiling; Mus; Muscle; Muscle Development; Muscle Fibers; Muscle Weakness; Muscular Atrophy; Mutant Strains Mice; Mutation; Myopathy; Names; Natural regeneration; Operative Surgical Procedures; Patients; Perinatal; Play; Production; Proliferating; Recovery; Regenerative Medicine; Reporting; Respiratory Muscles; Risk; Role; Route; Skeletal Muscle; Source; Stem cell transplant; Stem cells; Testing; Therapeutic; Thermogenesis; Tissues; Transplantation; abstracting; age related; blastocyst; cell motility; clinically significant; in utero; in vivo; individual patient; induced pluripotent stem cell; innovation; intraperitoneal; knockout gene; muscle transplantation; mutant; myogenesis; organ growth; organ regeneration; precursor cell; regenerative; repaired; response; sarcopenia; satellite cell; scale up; skeletal; stem cell population; stem cell technology; study characteristics

Abstract Chronic muscle diseases including Duchenne muscular dystrophy (DMD) and aging-related sarcopenia result in muscle weakness, loss of independence, and increased risk of death. In addition, traumatic muscle injury and loss due to accidents, surgery, and wartime injuries needs prolonged recovery. Skeletal muscle is a highly regenerative tissue in which satellite cells, a stem cell population for skeletal muscle, play essential roles in creating and repairing skeletal muscle. However, this potential ultimately fails with disease and aging. Autologous satellite cell transplantation is a potential approach to create and repair skeletal muscle fibers, but satellite cells are rare (a few % of all muscle nuclei) and often difficult to isolate. 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, efficient myogenic differentiation and the scale-up of myogenic differentiation remain elusive and must be developed further in order to generate effective cellular therapies. A key to the generation of human myogenic cells and skeletal muscle in a host animal is the selective knockout of genes in the blastocyst that are critical for organ development. Therefore, in this proposal, (1) we will determine to which extent mouse iPSC-derived limb skeletal muscle will be generated after injection of mouse iPSCs into Pax3 mutant mouse blastocysts, creating a niche in which stem cells can occupy and form skeletal muscle in the limb. This approach will provide evidence for the creation of entire skeletal muscle by mouse iPSCs in vivo. In addition, (2) we will generate a humanized skeletal muscle using Pax3 mutant mouse embryos via in utero injection of human iPSCs in combination with Pax3 mutant embryos and iPSCs. This concerted approach will help us to create iPSC-derived skeletal muscle and myogenic cells in vivo that can be transplanted into patients for a definitive cure of myopathic diseases and muscle injuries. In addition, the humanized skeletal muscle in mice will serve as an animal model to study the characteristics and regeneration characteristics of the human skeletal muscle diseases and responses to pharmacological agents and to provide a proof of concept for generating patient-derived cell and tissue sources for autologous muscle transplantation. .

抽象的 慢性肌肉疾病，包括杜兴（Duchenne）肌肉营养不良（DMD）和与衰老有关 在肌肉无力，失去独立性和死亡风险增加。另外，创伤性肌肉受伤 由于事故，手术和战时伤害而导致的损失需要长时间康复。骨骼肌是高度 再生组织，其中卫星细胞是一种用于骨骼肌的干细胞种群，在 创造和修复骨骼肌。但是，这种潜力最终会因疾病和衰老而失败。 自体卫星细胞移植是创建和修复骨骼肌纤维的潜在方法，但 卫星细胞很少见（所有肌肉核的百分之几），并且通常很难分离。患者衍生的诱导 多能干细胞（IPSC）是获得无限数量的肌原细胞的理想细胞来源 移植后逃脱免疫排斥。但是，有效的肌生成分化和扩大规模 肌源分化仍然难以捉摸，必须进一步发展才能产生有效的细胞 疗法。宿主动物中人类肌生成细胞和骨骼肌产生的关键是 胚泡中基因的选择性敲除对器官发育至关重要的基因。因此，在此提案中 （1）我们将在注射后确定小鼠IPSC衍生的肢体骨骼肌肉在多大程度上 将小鼠iPSC的摄入到PAX3突变体小鼠胚泡中，形成一个小众，其中干细胞可以占据和 在肢体中形成骨骼肌。这种方法将为创造整个骨骼肌的创造提供证据 由鼠标IPSC在体内。此外，（2）我们将使用PAX3突变体产生人源化的骨骼肌 小鼠胚胎通过子宫内注射人IPSC与PAX3突变胚和IPSC结合使用。 这种一致的方法将帮助我们在体内创建IPSC衍生的骨骼肌和肌原性细胞 可以将其移植到患者中，以确定肌病性疾病和肌肉损伤。此外， 小鼠中的人源化骨骼肌将作为动物模型，以研究特征和 人骨骼肌疾病的再生特征和对药理剂的反应 并提供用于生成自体肌肉的患者衍生细胞和组织源的概念证明 移植。 。