盘状结构域受体 2/膜联蛋白 A2/基质金属蛋白酶 13 环通过促进成纤维样滑膜细胞的迁移和侵袭促进关节炎中的关节破坏。
The discoidin domain receptor 2/annexin A2/matrix metalloproteinase 13 loop promotes joint destruction in arthritis through promoting migration and invasion of fibroblast-like synoviocytes.
机构信息
Fourth Military Medical University, Xi'an, China, and Ningxia Medical University, Yinchuan, China.
出版信息
Arthritis Rheumatol. 2014 Sep;66(9):2355-67. doi: 10.1002/art.38696.
OBJECTIVE
Discoidin domain receptor 2 (DDR-2)/matrix metalloproteinase (MMP) signaling is an important pathway involved in cartilage destruction in rheumatoid arthritis (RA). However, the molecular mechanisms of this pathway have not been clearly identified. This study was undertaken to screen key molecules involved in this pathway and evaluate their biologic functions in synovium invasion of RA.
METHODS
DDR-2-interacting proteins were examined in vitro by immunoprecipitation and mass spectrometry, and annexin A2 was acquired. The effects of annexin A2 on fibroblast-like synoviocyte (FLS) migration were evaluated using a Transwell invasion assay and an Erasion trace test. In Ddr2(-/-) mice with collagen-induced arthritis (CIA), hematoxylin and eosin (H&E) staining, immunohistochemical analysis, and Western blot analysis were used to assess expression of DDR-2, annexin A2, and MMP-13, as well as synovial hyperplasia. Rats with CIA were treated with lentivirus annexin A2 small interfering RNA (siRNA), and annexin A2 siRNA effects on joint damage were analyzed based upon arthritis index scores and results of micro-computed tomography and H&E staining. The differences between annexin A2 expression in clinical samples from RA and osteoarthritis patients were compared using Western blotting.
RESULTS
Annexin 2 was identified for the first time as a DDR-2 binding protein. It may be phosphorylated by phospho-DDR-2, leading to MMP-13 secretion. The annexin A2 phosphorylation level and MMP-13 expression level were decreased and collagen-induced joint damage greatly reduced in Ddr2(-/-) mice. Joint damage in rats with CIA was significantly ameliorated when annexin A2 was down-regulated. Annexin A2 expression and phosphorylation were elevated in human RA synovial tissue.
CONCLUSION
Annexin A2 is a key molecule in the DDR-2/annexin A2/MMP-13 loop, the activation of which contributes to joint destruction in RA, mainly through promoting invasion of FLS. Annexin A2 might therefore become a novel clinical target for RA treatment.
目的
盘状结构域受体 2（DDR-2）/基质金属蛋白酶（MMP）信号通路是类风湿关节炎（RA）软骨破坏的重要途径。然而，该途径的分子机制尚未明确。本研究旨在筛选该通路中的关键分子，并评估其在 RA 滑膜侵袭中的生物学功能。
方法
通过免疫沉淀和质谱分析检测 DDR-2 相互作用蛋白，并获得膜联蛋白 A2。通过 Transwell 侵袭实验和 Erase 痕迹实验评估膜联蛋白 A2 对成纤维样滑膜细胞（FLS）迁移的影响。在胶原诱导性关节炎（CIA）的 Ddr2(-/-) 小鼠中，采用苏木精和伊红（H&E）染色、免疫组织化学分析和 Western blot 分析评估 DDR-2、膜联蛋白 A2 和 MMP-13 的表达以及滑膜增生。用慢病毒膜联蛋白 A2 小干扰 RNA（siRNA）处理 CIA 大鼠，并根据关节炎指数评分以及 micro-CT 和 H&E 染色结果分析膜联蛋白 A2 siRNA 对关节损伤的影响。采用 Western blot 比较 RA 和骨关节炎患者临床样本中膜联蛋白 A2 的表达差异。
结果
首次鉴定出膜联蛋白 2 是 DDR-2 的结合蛋白。它可能被磷酸化的 DDR-2 磷酸化，导致 MMP-13 的分泌。在 Ddr2(-/-) 小鼠中，膜联蛋白 A2 的磷酸化水平和 MMP-13 的表达水平降低，胶原诱导的关节损伤大大减少。下调膜联蛋白 A2 后，CIA 大鼠的关节损伤明显改善。在人类 RA 滑膜组织中，膜联蛋白 A2 的表达和磷酸化水平升高。
结论
膜联蛋白 A2 是 DDR-2/膜联蛋白 A2/MMP-13 环中的关键分子，其激活促进了 RA 中的关节破坏，主要通过促进 FLS 的侵袭。因此，膜联蛋白 A2 可能成为治疗 RA 的新临床靶点。
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