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，往往导致功能恢复不良和 患者的生活质量受损。干细胞疗法前景看好；然而，它的临床应用 干细胞黏附有限和缺乏有效分化在很大程度上阻碍了移植。我们已经展示了 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 互动。因此，我们的具体目标是开发和优化新的候选类似物来促进人类 脂肪干细胞(HASC)的体外黏附和分化；细胞表面修饰 将技术引入基于HASC的治疗以促进周围神经再生；并调查相关 改善神经再生的机制。 目的：通过代谢糖工程(MGE)技术开发和优化新的类似物以 促进HASC的细胞黏附和细胞分化。我们将用以下方法优化细胞表面修饰 硫代糖类似物(ManNAc)，评估其效果，并对其进行全面表征以促进hASCs 黏附、增殖和分化。 目的：将MGE应用于HASC治疗，以促进周围神经再生。使用 优化的ManNAc类似物，我们将系统地评价糖工程hASCs对神经的影响 神经修复后再生，进一步优化治疗。 目标3： 检视 硫醇衍生的甘露醇类似物对神经的作用机制 再生 。 随着神经再生方面的预期改善， 我们将评估信号通路(例如，WNT / 骨髓间充质干细胞移植后的β。 -连环蛋白)由MGE调制 创新之处在于我们的假设，即用糖类似物来修饰干细胞表面的糖链性质 改善细胞存活和分化，我们的新和有效的技术，以及这些技术的新应用 技术在完全平移的神经修复模型中开发一种新的治疗方法。其意义在于 通过表面修饰来解决神经最具挑战性的一个方面的新型细胞疗法 修复临界大小的神经间隙的再生，以及潜在机制的预期发现 移植MGE‘-HASC可提高细胞的存活和分化能力。我们的技术和协议高度 可翻译到临床环境中。这个项目的成功将对以下内容产生直接的翻译影响 周围神经损伤需要手术修复的患者。ManNAc的临床研究表明 单次口服剂量高达6克的安全性，FDA已批准使用ManNAc治疗GNE肌病。 我们的研究将导致开发新的神经修复治疗策略，这将有助于 未来的临床干预，并根据新发现最大限度地发挥干细胞治疗的益处。

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.