基于单细胞测序技术筛选的源于MSCs的miR-17-92基因簇多重调控Kupffer细胞功能减轻移植肝损伤的机制研究

干细胞治疗在解决移植肝损伤这一难点问题方面展现良好前景，但因机制不明严重影响其临床应用。申请人通过单细胞测序技术发现源于MSCs的miR-17-92基因簇在调控Kupffer细胞功能，减轻移植肝损伤方面发挥重要作用。预实验结果结合生物信息学分析显示，该基因簇可以通过缝隙连接或外泌体传递到Kupffer细胞，作用于不同信号通路，减轻胞外诱捕网METs的直接杀伤作用，促进M1向M2极化转化，最终对移植肝产生综合保护作用。据此提出“源于MSCs的miR-17-92基因簇多重调控Kupffer细胞功能减轻移植肝损伤”的研究假设。本研究拟在前期基础上，通过体内外实验探讨miR-17-92基因簇的传递途径、作用的多种信号通路、对Kupffer细胞功能产生的多重调控作用以及对移植肝损伤的影响，阐明MSCs通过调控Kupffer细胞功能对移植肝产生保护作用的新机制，为干细胞的临床转化应用提供新的理论依据。

Stem cell therapy shows good prospects in solving the difficult problem of liver graft injury, but its clinical application is seriously affected by the unknown mechanism. The applicant discovered through single-cell sequencing technology that the miR-17-92 gene cluster derived from MSCs played an important role in regulating the functions of Kupffer cells and reducing the incidence of liver graft injury. Pre-experimental results combined with bioinformatics analysis show that miR-17-92 gene cluster can be transmitted to Kupffer cells through gap junctions or exosomes, acting on different signaling pathways, reducing the direct killing effect of extracellular trapping network METs, and promoting the polarization transformation of M1 to M2. Ultimately, it has comprehensive protective effects on the liver graft. Based on these results and analysis, the research hypothesis that the miR-17-92 gene cluster derived from MSCs multi-modulates the functions of Kupffer cells to reduce liver graft injury is proposed. This study intends to explore the transmission pathways of miR-17-92 gene cluster derived from MSCs to Kupffer cells through in vivo and in vitro experiments, the multiple signaling pathways that it acts on, the multiple regulatory effects on Kupffer cell functions and the impact on liver graft injury, so as to find out a new mechanism of MSCs protecting against liver graft injury by regulating the functions of Kupffer cells, and provide a new theoretical basis for the clinical application of stem cells.