过表达Rictor基因的供者Rapa-DCreg抑制小鼠肾移植排斥反应及机制研究

调节性树突状细胞（DCreg）治疗是目前最有潜力的抑制排斥反应策略之一，但药物诱导的DCreg在体内功能不稳定，且机制不详。本课题组前期发现雷帕霉素(Rapa)能有效诱导DCreg(Rapa-DCreg)，但Rapa仅抑制DCreg的mTORC1信号下游，未影响mTORC2信号下游，而DC在敲除 Rictor(mTORC2的核心成份)之后可加速移植排斥反应。故本项目采用药物诱导和基因修饰相结合的方法拟对供者Rapa-DCreg过表达Rictor产生调节功能更强、更稳定的Rictor-high-Rapa-DCreg，静脉输注到受体小鼠体内抑制肾移植排斥反应。运用现代分子生物学手段检测供体 DCreg的凋亡及其与受体DC的cross-dressing，双光子活体成像观察供体DCreg与受体DC和Treg的互动，多角度解析其作用机制。本研究为DCreg抑制排斥反应提供新的诱导方案和重要理论基础。

Regulatory dendritic cell (DCreg) therapy is currently one of the most promising strategies to suppress transplant rejection, but the function of drug-induced DCreg is not stable in vivo and the mechanisms are poorly understood. Our research group previously found that Rapamycin (Rapa) could effectively induce DCreg (Rapa-DCreg), however, Rapa only inhibited the downstream of mTORC1 in DCreg and it did not affect the downstream of mTORC2. Whereas, Rictor (a core component of mTORC2) specifically deficiency in DC can accelerate graft rejection. Therefore, this project will overexpress Rictor in Rapa-DCreg to generate regulatory functional stronger and stable “Rictor-high-Rapa-DCreg” through combining of drug induction and gene modification method. Intravenous infusion of Rictor-high-Rapa-DCreg into recipient can inhibit murine kidney transplant rejection. We will explore its mechanisms including detecting the apoptosis of donor DCreg and its cross-dressing with recipient DC by modern molecular biology techniques and monitoring the interactions of donor DCreg with recipient DC and Treg by intravital two-photon imaging. This project will provide a novel induction scheme and important theoretical basis for DCreg inhibiting rejection.