外泌体介导miR-23b分选拮抗Infliximab治疗类风湿性关节炎的调控机制探索

In the treatment of rheumatoid arthritis, nearly 50% of population has been demonstrated to possess the resistance of Infliximab, and meanwhile the drug could not block the damage of articular cartilage and bone tissue though it could control the inflammation process. For this phenomenon, the project explored the characteristics of origin and content of exosome and expression level of miR-23b in the synovial fluid of articular cavity through pharmacokinetic theory and multidisciplinary techniques. Further, the exosome-mediated miR-23b sorting in the fibroblast-like synoviocytes were systematically investigated to analyze the drug resistance mechanism, and key proteins and regulation mechanism were obtained by proteomics and gene editing strategies. Finally, the targets and mechanism of exosome harboring miR-23b on the damage of bone tissue would be studied through the interaction of exosome with osteoblasts and osteoclasts. Overall, the project provided a new insight into the role of exosome-mediated miR-23b sorting in the resistance of Infliximab during the treatment of rheumatoid arthritis. Meanwhile, it provided a theoretic basis for the Janus-type function of miR-23b in the rheumatoid arthritis, and thus it was of great significance for the development of targets in the treatment of rheumatoid arthritis.

在类风湿性关节炎治疗中，近50%的人群对英夫利昔单抗耐受，尽管炎症进程能够得到控制，却无法阻断关节软骨与骨组织的持续受损。针对这一现象，本项目利用药代动力学前沿理论结合多学科交叉技术手段，探究关节腔滑膜液中外泌体来源、含量及miR-23b表达水平的经时变化特征；阐明纤维样滑膜细胞通过外泌体介导miR-23b分选拮抗药物治疗的机制，并运用蛋白质组学、基因编辑等技术手段获得外泌体介导miR-23b分选的关键蛋白及调控机制；最后，通过内含miR-23b外泌体与成骨及破骨细胞的作用研究，阐明miR-23b作用骨组织相关细胞造成骨损伤的作用靶点及分子机制。本项目的实施，将为揭示外泌体介导miR-23b分选在英夫利昔单抗治疗类风湿性关节炎中的药物耐受机制提供新的研究思路，同时也为深入认识miR-23b这一“Janus型”分子的双重角色提供重要的理论支撑，对类风湿性关节炎治疗靶点的开发具有重要的意义。

在类风湿性关节炎治疗中，近50%的人群对英夫利昔单抗耐受，尽管炎症进程能够得到控制，但却无法阻断关节软骨与骨组织的持续受损。本项目以英夫利昔单抗在类风湿性关节炎临床应用中的药物耐受为研究对象，系统探索了外泌体介导miRNA分选与类风湿性关节炎治疗中药物耐受之间的相关性，通过miRNA表达谱及生物信息学分析，成功鉴定差异表达miRNA为miR-376b、miR-124a-2、miR-329及miR-190a，但尚未发现miR-23b与外泌体分选及英夫利昔单抗在类风湿性关节炎治疗中的药物耐受存在关联。幸运的是，我们成功挖掘到一类具有良好抗炎功能的寡核苷酸miR-30a，经确证，miR-30a能够显著下调Snail和Prlr基因的表达水平，其中Snail在类风湿性关节炎小鼠滑膜组织表达显著提升，与类风湿性关节炎的发生发展息息相关。在此基础上，以课题组前期构建的氟化修饰型树枝状高分子FP为载体，介导miR-30a的高效稳定递送，细胞水平上具有良好的炎症抑制能力，同时FP/miR-30a复合物能够有效抑制类风湿性关节炎小鼠发病进程，降低足部肿胀程度，恢复小鼠运动能力，并具有良好的生物相容性。综上，本项目旨在揭示外泌体介导miRNA分选在英夫利昔单抗治疗类风湿性关节炎中的药物耐受机制，系统挖掘了相关寡核苷酸分子的胞内作用靶点及拮抗类风湿性关节炎的分子机制，为揭示英夫利昔单抗治疗类风湿性关节炎中的药物耐受机制、类风湿性关节炎治疗靶点的开发及新型药物递送系统的设计提供了新的研究思路。