Examining the Therapeutic Potential of iPS cells in Duchenne Muscular Dystrophy

检查 iPS 细胞在杜氏肌营养不良症中的治疗潜力

• 批准号: 7808940
• 负责人: Rita C. R. Perlingeiro
• 金额: $ 99.97万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2012-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7808940
• 关键词: Adult; Autologous Transplantation; Cells; Clinic; Degenerative Disorder; Derivation procedure; Disease; Duchenne muscular dystrophy; Embryo; Engraftment; Ethical Issues; Fibroblasts; Future; Generations; Goals; Homologous Transplantation; Human; In Vitro; Injury; Knowledge; Methods; Mus; Muscle; Muscle Fibers; Muscle function; Muscular Dystrophies; Natural regeneration; Paraxial Mesoderm; Patients; Skeletal Muscle; Stem cells; Testing; Therapeutic; Translating; Transplantation; Wild Type Mouse; c-myc Genes; cell type; embryonic stem cell; human embryonic stem cell; improved; in vivo; induced pluripotent stem cell; injured; mdx mouse; novel; progenitor; public health relevance; regenerative; skeletal; stem

DESCRIPTION (provided by applicant): The recent breakthrough of direct reprogramming adult mouse and human fibroblasts toward a pluripotent state by introducing Oct3/4, Sox2, Klf4, and c-Myc, has brought new hope for the generation of patient- and disease- specific stem cells without the ethical issues associated with the derivation of human embryonic stem (ES) cells. However before induced pluripotent stem (iPS) cells can be applied for therapeutic applications, it is necessary to assess the ability of these cells to differentiate towards the desired cell type. For instance, skeletal myogenic progenitors are generated very inefficiently during in vitro differentiation of ES cells into embryoid bodies (EBs). This is due to the scarcity of paraxial mesoderm within EBs. We have recently demonstrated that Pax3 enables the generation of myogenic progenitors from differentiating ES cells that are endowed with the capacity to restore muscle function following their engraftment in mdx mice. Thus here we plan to examine the proof of principle that iPS cells may be used in the future for the treatment of DMD by combining the generation of iPS cells with our approach to derive myogenic progenitors by conditional expression of Pax7. Moreover, if one envisions translating iPS cells to the clinic, one has to demonstrate that functional myogenic progenitors can be successfully obtained from human ES cells. This will be assessed here by applying conditional expression of Pax7 to human ES cells with the goal to apply this knowledge to future studies involving human iPS cells obtained from patients with Duchene muscular dystrophy. PUBLIC HEALTH RELEVANCE: Embryonic stem (ES) cells and induced pluripotent stem (iPS) cells hold great promise for the treatment of degenerative diseases, however to date studies on their potential use in the treatment of muscular dystrophies have been hampered by the difficulty of differentiating ES cells into skeletal muscle progenitors. This application builds on a novel method we have developed to generate muscle progenitors from mouse ES cells. We have shown that such progenitors can be transplanted into normal injured, and dystrophic mice, where they contribute to muscle fiber regeneration, and improve muscle function after injury. In these studies, we will apply this approach to wild-type mouse iPS cells (Aim 1) as well as ex vivo genetically corrected dystrophic mouse iPS cells (Aim 2), thus assessing proof-of-principle to whether these cells are endowed with in vivo regenerative potential. In Aim 3, we will investigate the mechanisms controlling muscle differentiation in human ES cells with the goal to apply this knowledge to future studies involving human iPS cells obtained from patients with Duchenne muscular dystrophy.

描述（由申请人提供）：通过引入Oct3/4，Sox2，KLF4和C-Myc，直接重编程成年小鼠和人类成纤维细胞的最新突破使人对患者和疾病特异性干细胞的产生带来了新的希望，而没有与人类胚胎干细胞（ES）细胞产生的诚实问题。但是，在诱导多能干（IPS）细胞可以用于治疗应用之前，有必要评估这些细胞朝向所需的细胞类型的能力。例如，在将ES细胞分化为胚胎体（EBS）期间，骨骼肌祖细胞非常低效地产生。这是由于EBS中近去中胚层的稀缺性。我们最近证明，PAX3使肌源祖细胞与分化ES细胞的产生，这些细胞具有在MDX小鼠中植入后恢复肌肉功能的能力。因此，在这里，我们计划检查原理证据，即将来可以将IPS细胞用于处理DMD，通过将IPS细胞的产生与通过PAX7的条件表达来得出肌源祖细胞的方法来治疗DMD。此外，如果人们设想将IPS细胞转换为诊所，则必须证明可以成功地从人ES细胞中获得功能性肌源祖细胞。此处将通过将PAX7的条件表达应用于人类ES细胞的条件表达来进行评估，目的是将这些知识应用于未来的研究中，该研究涉及从Duchene肌肉营养不良患者获得的人IPS细胞。 公共卫生相关性：胚胎干细胞和诱导的多能干（IPS）细胞对治疗退行性疾病具有很大的希望，但是，迄今为止，关于它们在治疗肌肉营养不良治疗中的潜在使用的研究已受到将ES细胞在骨骼肌中的难度受到阻碍。该应用以一种新的方法为基础，我们已经开发出来从小鼠ES细胞中产生肌肉祖细胞。我们已经表明，这种祖细胞可以被移植到正常受伤的和营养不良的小鼠中，它们会导致肌肉纤维再生，并在受伤后改善肌肉功能。在这些研究中，我们将将这种方法应用于野生型小鼠IPS细胞（AIM 1）以及离体遗传校正的营养不良小鼠IPS细胞（AIM 2），从而评估了这些细胞是否赋予这些细胞是否具有体内再生势。在AIM 3中，我们将研究控制人类ES细胞中肌肉分化的机制，其目标是将这些知识应用于未来的研究，涉及从Duchenne肌肉营养不良患者获得的人IPS细胞。