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

项目摘要 杜氏肌营养不良症是一种遗传性疾病，由一种称为肌营养不良蛋白的蛋白质缺陷引起。 肌营养不良蛋白的功能是为肌细胞提供结构和机械稳定性; 杜氏肌营养不良症中的功能性肌营养不良蛋白导致进行性肌肉变性，虚弱， 以及由于心肺衰竭而显著缩短的预期寿命。无药可救。作为一个潜在 治愈性治疗，通过肌肉干细胞移植恢复肌营养不良蛋白正在积极探索中， 但是这些努力的临床转化受到无法产生大量肌干的阻碍 在不影响其治疗效力的情况下。此外，拓展也极具挑战性 肌肉干细胞来源于培养皿中的营养不良肌肉，这是由于其固有的功能缺陷。这进一步 将策略限制在体外对患者来源的肌肉干细胞进行基因校正，在培养皿中培养它们， 自体移植功能性肌肉干细胞。在这个多样性补充申请中， 我们将开发一种Notch调节肌肉干细胞培养平台， 大量治疗有效的肌肉干细胞。这项研究加强和扩大了 父R 01项目，通过工程化水凝胶平台来支持营养不良的肌肉细胞扩增， 治疗应用。为了实现这一目标，Aim 1将设计一种Notch调节水凝胶， 离体营养不良的肌肉干细胞。目的2：探讨抑制p38α/β丝裂原活化的 维持在Notch调节水凝胶内的营养不良肌肉干细胞的蛋白激酶（MAPK）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，用于安全治疗杜氏肌营养不良症，在这种技术也将使前 在重新引入以恢复之前，对患者体外的肌肉干细胞进行体内基因校正 抗肌萎缩蛋白。

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.