Genome-wide microRNA screening reveals miR-582-5p as a mesenchymal stem cell-specific microRNA in subchondral bone of the human knee joint
Genome-wide microRNA screening reveals miR-582-5p as a mesenchymal stem cell-specific microRNA in subchondral bone of the human knee joint
复制标题
全基因组 microRNA 筛查揭示 miR-582-5p 是人类膝关节软骨下骨中的间充质干细胞特异性 microRNA
DOI：
10.1002/jcp.28751
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发表时间：
2019-12-01
影响因子：
5.6
通讯作者：
Jin, Hongting
中科院分区：
文献类型：
--
作者：
Wang, Pinger;Dong, Rui;Jin, Hongting
新的证据表明，microRNAs(MiRNAs)可能在病理上参与了骨关节炎(OA)。软骨下骨硬化症(SCB)是膝关节骨关节炎(KOA)发生和发展的重要原因。在本研究中，我们旨在筛选KOA的miRNA生物标志物，并研究这些miRNA是否调节间充质干细胞(MSCs)的分化潜能，从而促进SCB的发生。我们通过微阵列表达谱检测在KOA患者(n=5)的血液样本中鉴定出48个miRNAs。在大样本验证后，我们证实hsa-miR-582-5p和hsa-miR-424-5p与SCB硬化的病理相关。利用在线数据库对目的基因进行预测和通路分析，表明这两个候选miRNAs与干细胞多能性途径和骨性关节炎的病理机制密切相关。令人惊讶的是，MMU-miR-582-5p(hsa-miR-582-5p的同源物)在成骨分化中下调，在间充质祖细胞C3H10T1/2成脂分化中上调，而MMU-mir-322-5p(hsa-miR-424-5p的同源物)在体外研究中没有变化。补充MMU-miR-582-5p模拟抑制C3H10T1/2细胞成骨和成脂分化，而沉默内源性MMU-miR-582-5p促进成骨和抑制成脂分化。进一步的机制研究表明，MMU-miR-582-5p直接靶向Runx2。Runx2 3‘非翻译区可能存在的MMU-miR-582-5P结合位点的突变可以取消3’UTR-荧光素酶结构对补充MMU-miR-582-5P的反应。总的来说，我们的数据提示miR-582-5p是骨性关节炎的一个重要的生物标志物，并且能够通过靶向Runx2来调节MSCs的成骨和成脂分化。该研究还提示miR-582-5p可能在人膝关节骨性关节炎的SCB硬化中起关键作用。
Emerging evidence suggests that microRNAs (miRNAs) may be pathologically involved in osteoarthritis (OA). Subchondral bone (SCB) sclerosis is accounted for the knee osteoarthritis (KOA) development and progression. In this study, we aimed to screen the miRNA biomarkers of KOA and investigated whether these miRNAs regulate the differentiation potential of mesenchymal stem cells (MSCs) and thus contributing to SCB. We identified 48 miRNAs in the blood samples in KOA patients (n=5) through microarray expression profiling detection. After validation with larger sample number, we confirmed hsa-miR-582-5p and hsa-miR-424-5p were associated with the pathology of SCB sclerosis. Target genes prediction and pathway analysis were implemented with online databases, indicating these two candidate miRNAs were closely related to the pathways of pluripotency of stem cells and pathology of OA. Surprisingly, mmu-miR-582-5p (homology of hsa-miR-582-5p) was downregulated in osteogenic differentiation and upregulated in adipogenic differentiation of mesenchymal progenitor C3H10T1/2 cells, whereas mmu-mir-322-5p (homology of hsa-miR-424-5p) showed no change through the in vitro study. Supplementing mmu-miR-582-5p mimics blocked osteogenic and induced adipogenic differentiation of C3H10T1/2 cells, whereas silencing of the endogenous mmu-miR-582-5p enhanced osteogenic and repressed adipogenic differentiation. Further mechanism studies showed that mmu-miR-582-5p was directly targeted to Runx2. Mutation of putative mmu-miR-582-5p binding sites in Runx2 3 ' untranslated region (3 ' UTR) could abolish the response of the 3 ' UTR-luciferase construct to mmu-miR-582-5p supplementation. Generally speaking, our data suggest that miR-582-5p is an important biomarker of KOA and is able to regulate osteogenic and adipogenic differentiation of MSCs via targeting Runx2. The study also suggests that miR-582-5p may play a crucial role in SCB sclerosis of human KOA.