Stem Cell Surface Modification to Promote Nerve Regeneration

项目摘要 周围神经损伤，特别是临界大小的神经间隙损伤，通常导致功能恢复不良， 患者的生活质量受损。干细胞治疗具有重大的前景;然而，其临床应用 由于干细胞粘附有限和缺乏有效的分化而受到很大的阻碍。我们已经表明 我们的干细胞表面修饰技术能够深刻地影响特定的细胞-细胞和细胞-基质 交互.因此，我们的具体目标是开发和优化新的候选类似物，以促进人类 脂肪干细胞（hASC）的体外粘附和分化;将细胞表面修饰 技术用于基于hASC的治疗以改善周围神经再生;并研究相关的 改善神经再生的潜在机制。 目的1：通过代谢糖工程（MGE）技术开发和优化新型类似物， 促进hASC体外细胞粘附和细胞分化。我们将优化细胞表面修饰， 巯基化糖类似物（ManNAc），评估其作用，并彻底表征其促进hASC 粘附、增殖和分化。 目的2：将MGE纳入基于hASC的疗法中以改善周围神经再生。与 优化的ManNAc类似物，我们将系统地评估糖工程化的hASCs对神经元的作用。 神经修复后的再生，并进一步优化治疗。 目标3： 审查 巯基衍生ManNAc类似物有助于神经的机制 再生 . 随着神经再生的预期改善， 我们将评估信号通路（例如，Wnt / MGE'ed艾德hASC移植后β。 - 连环蛋白） 创新在于我们的假设，即用糖类似物修饰干细胞表面聚糖的性质， 改善细胞存活和分化，我们的新的和有效的技术，以及这些新的应用 技术在完全平移神经修复模型中开发新的治疗方法。其意义在于 一种新型的基于细胞的表面修饰疗法，以解决神经系统最具挑战性的方面之一。 再生的临界大小的神经间隙修复，以及预期的发现的机制， 通过移植的MGE'ed艾德hASC改善存活和分化。我们的技术和协议 可移植到临床环境中。该项目的成功将对以下方面产生直接的翻译影响： 需要手术修复的周围神经损伤患者。ManNAc的临床研究表明， 单次口服剂量高达6 g的安全性，FDA已批准使用ManNAc治疗GNE肌病。 我们的研究将导致神经修复的新治疗策略的发展， 未来的临床干预措施，并最大限度地发挥干细胞治疗的好处的基础上的新发现。

Project Summary Peripheral nerve injury, especially critical-sized nerve gap injury, often results in poor recovery of function and impaired quality of life for the patient. Stem cell therapy holds significant promise; however, its clinical application has been largely hampered by limited stem cell adhesion and the lack of efficient differentiation. We have shown that our stem cell surface modification technique is able to profoundly influence specific cell-cell and cell-matrix interactions. Therefore, our specific aims are to develop and optimize novel candidate analogs to promote human adipose stem cell (hASC) adhesion and differentiation in vitro; to incorporate the cell surface modification technique into hASC-based therapies to improve peripheral nerve regeneration; and to investigate related mechanisms underlying improved nerve regeneration. Aim1: To develop and optimize novel analogs by metabolic glycoengineering (MGE) technology to promote hASC's cell adhesion and cell differentiation in vitro. We will optimize the cell surface modification with thiolated sugar analogs (ManNAc), evaluate the effects, and thoroughly characterize them to promote hASCs adhesion, proliferation, and differentiation. Aim2: To incorporate MGE into hASC-based therapies to improve peripheral nerve regeneration. With optimized ManNAc analogs, we will systemically evaluate the effect of glycoengineered hASCs on nerve regeneration after nerve repair and further optimize the therapy. Aim 3: To examine the mechanism by which thiol-derivatized ManNAc analogs contribute to nerve regeneration . With expected improvements in nerve regeneration, we will evaluate signaling pathways (e.g., Wnt / β after MGE'ed hASC transplantation. -catenin) modulated by MGE The innovation lies in our hypothesis to modify stem cell surface glycan properties with sugar analogs to improve cell survival and differentiation, our novel and effective technology, and the new application of these technologies in a fully translational nerve repair model to develop a novel treatment. The significance lies in the novel cell-based therapy with surface modification to address one of the most challenging aspects of nerve regeneration for critical-sized nerve gap repair, and the expected discovery of the mechanism underlying improved survival and differentiation by transplanted MGE'ed hASC. Our technology and protocols are highly translatable to the clinical environment. Success in this project will have direct translational implications for patients with peripheral nerve trauma requiring surgical repair. The clinical study of ManNAc has demonstrated the safety of single oral doses up to 6 g, and the FDA has approved the use of ManNAc to treat GNE Myopathy. Our study will lead to the development of novel therapeutic strategies for nerve repair that can contribute to future clinical interventions and maximize the benefits of stem cell therapy based on the new findings.