Liu Chong, Liang Tuo, Zhang Zide, Chen Jiarui, Xue Jang, Zhan Xinli, Ren Liang
Spine and Osteopathy Ward, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, P. R. China.
Guangxi Medical University, Nanning, 530021, P. R. China.
Cell Death Discov. 2022 May 23;8(1):269. doi: 10.1038/s41420-022-00900-1.
Pathological osteogenesis and inflammation possess critical significance in ankylosing spondylitis (AS). The current study aimed to elucidate the mechanisms regarding extracellular vesicle (EV)-packaged microRNA-22-3p (miR-22-3p) from M2 macrophages in the osteogenic differentiation of mesenchymal stem cells (MSCs) in AS. EVs were initially isolated from M2 macrophages, which had been treated with either restored or depleted miR-22-3p. AS-BMSCs were subsequently treated with M2 macrophage-derived EVs to detect osteogenic differentiation in BMSCs using gain- or loss-of-function experiments. The binding affinity among miR-22-3p, period circadian protein 2 (PER2), and Wnt7b was identified. Finally, AS mouse models were established for testing the effects of M2-EV-miR-22-3p on the bone metastatic microenvironment in vivo. miR-22-3p from M2 macrophages could be transferred into BMSCs via EVs, which promoted the osteogenic differentiation of AS-BMSCs. miR-22-3p inhibited PER2, while PER2 blocked the Wnt/β-catenin signaling pathway via Wnt7b inhibition. M2-EV-shuttled miR-22-3p facilitated alkaline phosphatase activity and extracellular matrix mineralization via PER2-regulated Wnt/β-catenin axis, stimulating the BMSC osteogenic differentiation. Taken together, these findings demonstrate that miR-22-3p in M2 macrophage-released EVs downregulates PER2 to facilitate the osteogenesis of MSCs via Wnt/β-catenin axis.
病理性骨生成和炎症在强直性脊柱炎（AS）中具有关键意义。本研究旨在阐明来自M2巨噬细胞的细胞外囊泡（EV）包裹的微小RNA-22-3p（miR-22-3p）在AS中间充质干细胞（MSC）成骨分化中的机制。首先从用恢复或缺失miR-22-3p处理过的M2巨噬细胞中分离出EV。随后用M2巨噬细胞衍生的EV处理AS-BMSC，通过功能获得或功能缺失实验检测BMSC中的成骨分化。确定了miR-22-3p、周期昼夜节律蛋白2（PER2）和Wnt7b之间的结合亲和力。最后，建立AS小鼠模型以测试M2-EV-miR-22-3p在体内对骨转移微环境的影响。来自M2巨噬细胞的miR-22-3p可通过EV转移到BMSC中，促进AS-BMSC的成骨分化。miR-22-3p抑制PER2，而PER2通过抑制Wnt7b阻断Wnt/β-连环蛋白信号通路。M2-EV穿梭的miR-22-3p通过PER2调节的Wnt/β-连环蛋白轴促进碱性磷酸酶活性和细胞外基质矿化，刺激BMSC成骨分化。综上所述，这些发现表明M2巨噬细胞释放的EV中的miR-22-3p通过Wnt/β-连环蛋白轴下调PER2以促进MSC的成骨作用。
