本研究前期发现：携带α-突触核蛋白（α-syn）的帕金森病患者血浆外泌体注入小鼠纹状体后可被小胶质细胞高度选择性摄取（而非神经元和星形胶质细胞），黑质TLR2表达增加。外泌体处理小胶质细胞后，其自噬明显受抑制，磷酸化的AKT和mTOR的表达明显增加。结合文献我们推测TLR2-AKT-mTOR通路在小胶质细胞介导的外泌体α-syn传播中可能发挥了重要作用。本项目将采用超高分辨活细胞成像技术，观察小胶质细胞吞噬外泌体的动态过程，进一步研究小胶质细胞在外泌体α-syn传播中的呈递作用；采用基因编辑及特异性抗体竞争结合等技术，分别在细胞和小鼠模型中抑制小胶质细胞分泌外泌体、阻断α-syn与TLR2结合，从而阐明上述通路在α-syn的识别内化和释放调节中的作用及机制；最后通过减少小胶质细胞激活、干预AKT-mTOR自噬活性进而减少外泌体α-syn的传播来干预帕金森病的病理进程，为其治疗提供新的视角。

Previous results of this study showed that PD exosomes carrying α-syn oligomers were stereotaxically injected into the unilateral striatum of mice brains, which could be highly selective uptake by microglia (rather than neurons and astrocytes), and resulted in an increase in the TLR2 expression of substantia nigra. After the treatment of microglia by PD exosomes, autophagy was significantly inhibited, and the expression of phosphorylated AKT and mTOR was significantly increased. Based on the literature, we speculate that TLR2-AKT-mTOR pathway may play an important role in microglia-mediated exosomal α-syn transmission. In this project, the dynamic process of microglia phagocytosis of exosomes will be observed by using the new technique of ultra-high resolution living cell imaging, and will further investigate the role of microglia in the exosomal α-syn transmission; gene editing and specific antibody binding techniques were used to inhibit exosomes secretion of microglia and block the binding of a-syn and TLK2 in cell and mouse models, respectively; so as to elucidate the role and mechanism of TLR2-AKT-mTOR pathway in the regulation of the recognition and release of α-syn; finally, the propagation of exosomal a-syn was reduced by repressing the activation of microglial cells and interfering with autophagy activity, which providing a new perspective for the study of PD neuroprotection.