Brain Recovery after Cardiac Arrest with Metabolic Glycoengineered Stem Cells

项目摘要 心脏骤停(CA)每年的发病率为35.98万。在CA的幸存者中，脑损伤是最大的 功能恢复障碍。目前，无论是药物干预还是治疗 亚低温可逆转CA所致的神经损伤。干细胞疗法在治疗癌症方面前景看好 脑损伤后的神经元修复。然而，损伤部位的生存能力和整合力较差，缺乏效率 分化为所需的细胞类型阻碍了临床应用。E 正在合并 代谢性糖工程 然而，通过修饰表面多聚糖(MGE)技术在体外影响细胞黏附和分化， 尚未在干细胞治疗的背景下进行研究。因此，这项建议的总体目标是 将MGE应用于基于细胞的治疗，以改善移植后的细胞黏附和生存能力，并增强 CA后脑缺血损伤神经元修复的治疗效果。具体目标是： 目的：通过我们的新的MGE技术，我们假设一种新的基于多糖的干预能够 促进人神经干细胞(HNSCs)体外神经分化和细胞黏附。我们将发展和 优化具有更长烷链的新型硫代化ManNAc类似物，Ac5ManNPropT和Ac5ManNButT，这两种化合物是 预计在体外可增加硫醇可及性，促进hNSCs的细胞黏附和神经分化。 AIM2：通过优化的ManNAc类似物，我们假设经处理的hNSCs将促进存活， 移植的hNSCs在体内的分布和分化。我们将评估糖工程的效果 HNSCs对CA后的功能结果的影响，并优化这一基于细胞的治疗。 目标3：随着CA术后预后的预期改善，我们假设细胞的成功- 基础干预是由于提高了移植的糖工程人神经干细胞的存活率和分化能力。我们 将探讨糖工程hNSC移植后的细胞相互作用和分子机制 CA通过Wnt/β-catenin信号通路。 MGE技术与干细胞治疗相结合对脑损伤修复的意义 CA后，以细胞为基础的治疗向临床转化的优化，以及预期的发现 糖工程神经干细胞移植后改善存活和分化的机制。这个 创新之处在于我们的创新假设，即通过MGE技术修改干细胞表面特性，以 改善细胞存活和分化，我们新的低成本的表面修饰有效的MGE方法 HNSCs的糖链，以及我们在重要疾病体内模型中使用MGE技术开发新的 基于细胞的治疗性干预。我们的研究将导致开发新的治疗策略 为未来的临床干预修复脑损伤并最大限度地发挥MGE和干细胞治疗的好处 根据新的发现。糖类似物分子在再生医学和干细胞治疗中的应用 将有助于改进基于细胞的治疗，以修复因CA、中风和创伤或神经退行性疾病而造成的脑损伤 疾病，并具有巨大的潜力，提供深刻的医学进步。

Project Summary Cardiac arrest (CA) has an incidence of 359,800 annually. Among survivors of CA, brain injury is the biggest impediment to functional recovery. Currently, neither pharmacological intervention nor therapeutic hypothermia can reverse the neural injury caused by CA. Stem cell therapy holds significant promise in the neuronal repair after brain injury. However, poor viability and integration at the site of injury and lack of efficient differentiation into the desired cell types hinder clinical applications. E merging metabolic glycoengineering (MGE) technology by modification of surface glycans impacts cell adhesion and differentiation in vitro, however, has not been investigated in the context of stem cell therapy. Therefore, the overall aim of this proposal is to apply MGE to cell-based therapies to improve cell adhesion and viability after transplantation and enhance the treatment efficacy to repair damaged neurons in ischemia brain after CA. The specific aims are: Aim1: With our novel MGE technique, we hypothesize that a novel glycan-based intervention is able to promote human neural stem cell (hNSCs) neural differentiation and cell adhesion in vitro. We will develop and optimize novel thiolated ManNAc analogs with longer alkyl chains, Ac5ManNPropT and Ac5ManNButT, that are predicted to increase thiol accessibility and promote hNSCs cell adhesion and neural differentiation in vitro. Aim2: With optimized ManNAc analogs, we hypothesize that treated hNSCs will promote the survival, distribution, and differentiation of transplanted hNSCs in vivo. We will evaluate the effect of glycoengineered hNSCs on functional outcome after CA and optimize this cell-based therapy. Aim 3: With expected improvement in outcome after CA, we hypothesize that the success of the cell- based intervention is due to improved survival and differentiation of transplanted glycoengineered hNSCs. We will explore cellular interactions and molecular mechanisms after glycoengineered hNSC transplantation after CA through Wnt/β-catenin signaling pathways. The Significance lies in the combination of the MGE technique and stem cell therapy for repairing brain injury post-CA, optimization of cell-based therapy towards clinical translation, and the expected discovery of the mechanism underlying improved survival and differentiation after glycoengineered NSC transplantation. The innovation lies in our innovative hypothesis to modify stem cell surface properties by MGE technology to improve cell survival and differentiation, our novel and effective MGE method with low cost for modifying surface glycans of hNSCs, and our use of the MGE technique in important disease in vivo model to develop novel therapeutic cell-based intervention. Our study will lead to the development of novel therapeutic strategies to repair brain injury towards future clinical interventions and maximize the benefits of MGE and stem cell therapy based on the new findings. The use of sugar analog molecules for regenerative medicine and stem cell therapies will help improve cells based therapy to repair brain injury due to CA, stroke, and trauma, or neurodegenerative diseases, and have tremendous potential to provide a profound medical advance.