MSCs源exosomes负载α-突触核蛋白相关rno-miR-7对PD模型的联合作用研究

The objective of the project is to investigate the potential co-application of stem cell transplantation and delivery gene therapy in the treatment of Parkinson's disease (PD) by introducing cellular transport vesicles-exosomes into PD model, since stem cell-derived exosomes can play dual function as stem cells information and gene therapy deliveries. The repair effects of exosomes secreted from mesenchymal stem cells (MSCs) of different sources on damaged nerve in PD model were firstly studied by parallel three-dimensional perfusion bioreactor and MSCs-derived exosomes with high repair ability was screened. Secondly, the technique of exosomes transfected with micro RNA was developed and MSCs-exosomes-miRNA complex was constructed. Our study have improved and developed the exosomes labeling technology by quantum dot to facilitate the long-term stable in vivo monitoring of exosomes. Then PD cell model with Thy-1 overexpression and Thy-1transgenic PD animal models were employed in the project to study the effects and mechanism of MSCs-exosomes-miRNA complexes on PD treatment in vivo and in vitro. We proposed for the first time a new attempt by intranasal administration of MSCs-exosomes-miRNA complex for PD therapy. The project also explored new ideas for PD therapy, using the micro RNA delivered by MSCs-derived exosomes and its mechanism. Our results may put forward new approaches to PD treatment by vesicle transport and gene-targeted delivery, which has significant clinical applications and social values.

近来干细胞分泌的细胞囊泡exosomes在神经损伤修复中的作用受到关注,而其来源于机体自然成分、良好的RNA转运能力及体液中稳定存在的特点,更显示出其在基因治疗中的巨大优势。项目拟发挥间充质干细胞（MSCs）源-exosomes作为干细胞信息携带者和基因治疗承载体的双重功能，以Thy-1 过表达帕金森病(PD)细胞模型及Thy-1 转基因PD动物模型为研究对象，从体内外综合研究MSCs源exosomes负载α-突触核蛋白相关rno-miR-7植入在PD治疗中的联合作用及机制。项目开辟了一个PD治疗的新理念，寻求到一条全新的治疗途径，通过探讨其可行性及技术可操作性，为今后发展PD联合治疗模式奠定基础理论及实验基础，具有重要的临床应用价值与社会意义。

项目评估了不同充质干细胞来源外泌体的神经修复功能，从中筛选出了高效损伤修复能力的外泌体；以量子点为荧光探针，构建了灵敏高效且靶向性强的外泌体标记技术；成功实现了MSCs-外泌体转染PD相关治疗基因miRNA；基于DA能功能神经网络研究了MSCs-外泌体对损伤DA能神经元的修复作用及作用机制；基于Thy-1过表达帕金森细胞模型及Thy-1转基因PD动物模型，体内外综合评价了MSCs-外泌体-miRNA复合体对PD的联合修复作用。项目开辟了一个PD治疗的新理念，通过探讨其可行性及技术可操作性，为今后发展PD联合治疗模式奠定理论基础及实验基础。