Notch-Modulatory Hydrogel for Dystrophic Muscle Stem Cell Rejuvenation and Expansion

项目摘要 Duchenne肌营养不良症是一种遗传性疾病，由一种名为dystrophin的蛋白质缺陷引起。 肌营养不良蛋白的功能是为肌肉细胞提供结构和机械稳定性；以及缺乏 Duchenne肌营养不良症的功能性肌营养不良蛋白导致进行性肌肉退化，无力， 以及由于心肺衰竭而导致的预期寿命显著缩短。没有治愈的方法。作为一种潜力 治疗，通过肌肉干细胞移植恢复肌营养不良蛋白正在积极探索， 但是，由于无法产生大量的肌干，这些努力的临床翻译受到阻碍。 在不影响其治疗效力的情况下，培养皿中的细胞。此外，扩张也非常具有挑战性。 肌肉干细胞来源于营养不良的肌肉，由于其固有的功能缺陷。这进一步 限制体外对患者来源的肌肉干细胞进行基因纠正的策略，将它们培养在培养皿中，以及 以自体的方式移植有功能的肌肉干细胞。在该多样性补充应用中， 我们将开发一种Notch调节型肌肉干细胞培养平台，使大鼠 大量具有治疗能力的肌肉干细胞。这项研究加强和扩大了研究的范围 通过设计水凝胶平台来支持营养不良肌肉细胞的细胞扩增，从而为R01项目提供母项目 治疗应用。为了实现这一目标，目标一号将设计一种Notch-调制型水凝胶来恢复活力 营养不良的肌肉干细胞体外培养。目标2将确定抑制p38α/β丝裂原激活的效果 营养不良肌肉干细胞在Notch调控水凝胶中的蛋白激酶活性 长期的。这项研究的结果将开辟一条通往个性化(自体)肌肉的道路 干细胞疗法用于安全治疗Duchenne肌营养不良症，这种技术也将使EX 对患者体外肌肉干细胞进行体内基因矫正，然后重新引入修复 营养不良蛋白。

PROJECT ABSTRACT Duchenne muscular dystrophy is a genetic disease caused by defects in a protein known as dystrophin. Dystrophin functions to provide structural and mechanical stability to the muscle cells; and the absence of functional dystrophin in Duchenne muscular dystrophy leads to progressive muscle degeneration, weakness, and a significantly reduced life expectancy due to cardiorespiratory failure. There is no cure. As a potential curative therapy, restoration of dystrophin through muscle stem cell transplantation is actively being explored, but the clinical translation of these efforts is hampered by the inability to generate large quantities of muscle stem cells in dishes without compromising their therapeutic potency. In addition, it is also very challenging to expand muscle stem cells derived from dystrophic muscles in dishes due to their inherent functional deficits. This further limits strategies to gene correct patient-derived muscle stem cells outside the body, grow them in a dish, and transplant the functional muscle stem cells in an autologous manner. In this Diversity Supplement application, we will develop a Notch-modulatory muscle stem cell culture platform that enables the generation of large quantities of therapeutically potent muscle stem cells. This research strengthens and expands the scope of the parent R01 project by engineering a hydrogel platform to support dystrophic muscle cell expansion for cell therapy applications. Towards this goal, Aim 1 will engineer a Notch-modulatory hydrogel for rejuvenating dystrophic muscle stem cells ex vivo. Aim 2 will determine the effects of suppressing p38α/β mitogen-activated protein kinase (MAPK) activity of dystrophic muscle stem cells maintained within the Notch-modulatory hydrogel long-term. The outcomes of this research will trailblaze a pathway towards personalized (autologous) muscle stem cell therapy for safely treating Duchenne muscular dystrophy, where such technologies will also enable ex vivo gene correction on the muscle stem cells outside the patient's body before being re-introduced to restore dystrophin.