Aberrant Fluid Shear Stress Contributes to Articular Cartilage Pathogenesis via Epigenetic Regulation of ZBTB20 by H3K4me3.
Aberrant Fluid Shear Stress Contributes to Articular Cartilage Pathogenesis via Epigenetic Regulation of ZBTB20 by H3K4me3.
复制标题
异常的流体剪切应力通过 H3K4me3 对 ZBTB20 的表观遗传调控促进关节软骨发病
DOI：
10.2147/jir.s339382
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发表时间：
2021
影响因子：
4.5
通讯作者：
Xia L
中科院分区：
文献类型：
--
作者：
Jin Y;Li Z;Wu Y;Li H;Liu Z;Liu L;Ouyang N;Zhou T;Fang B;Xia L
目的骨关节炎是人类的常见病，以严重的炎症、软骨退化和软骨下骨破坏为特征。然而，目前的治疗方法仅限于缓解疼痛或关节置换，还没有发现有效的治疗方法来改善退行性变化。目前，已有多种证据表明，机械刺激异常与关节发病密切相关，但其具体机制尚不清楚。在本研究中，我们决定研究过高的流体剪应力(FSS)对原代软骨细胞的影响及其潜在的表观遗传学机制。材料与方法采用菲罗兰染色和EDU染色评价细胞形态和活力。采用定量聚合酶链式反应、蛋白印迹和免疫荧光染色检测基因的表达水平。通过RNA测序和切割标签测序进行机制研究。在体内，我们采用单侧前交叉咬合(UAC)小鼠模型，研究H3K4me3和ZBTB20在异常力相关软骨发病机制中的表达。结果FSS对软骨细胞形态有明显的破坏作用，细胞活力明显降低。异常的FSS可诱导显著的炎症介质产生，导致软骨的退化和降解。深入的机制研究表明，FSS使H3K4me3的表达上调10倍以上，H3K4me3对软骨的调控作用是通过直接靶向ZBTB20获得的。此外，在高FSS诱导的骨性关节炎发病机制中，Wnt信号被强烈激活，而ZBTB20对软骨细胞的负面影响也是通过激活Wnt信号途径实现的。此外，用MM-102药理抑制H3K4me3活化或用Wnt途径抑制剂LF3处理均可有效减轻FSS对软骨细胞的破坏作用。在活体UAC小鼠模型中，证实了H3K4me3和ZBTB20在反常力诱导软骨发病机制中的失调。结论通过体外FSS模型和体内UAC模型的结合，首次发现KMT2B-H3K4me3-ZBTB20轴参与了FSS诱导的软骨畸形的发病机制，为今后的表观遗传学治疗提供了依据。
Purpose Osteoarthritis (OA) is a common disease for human beings, characterized by severe inflammation, cartilage degradation, and subchondral bone destruction. However, current therapies are limited to relieving pain or joint replacement and no effective treatment methods have been discovered to improve degenerative changes. Currently, a variety of evidences have indicated that aberrant mechanical stimuli is closely associated with articular joint pathogenesis, while the detailed underlying mechanism remains unelucidated. In the present study, we determined to investigate the impact of excessive high fluid shear stress (FSS) on primary chondrocytes and the underlying epigenetic mechanisms. Materials and Methods Phalloidin staining and EdU staining were used to evaluate cell morphology and viability. The mRNA level and protein level of genes were determined by qPCR, Western blot assay, and immunofluorescence staining. Mechanistic investigation was performed through RNA-sequencing and CUT&Tag sequencing. In vivo, we adopted unilateral anterior crossbites (UAC) mice model to investigate the expression of H3K4me3 and ZBTB20 in aberrant force-related cartilage pathogenesis. Results The results demonstrated that FSS greatly disrupts cell morphology and significantly decreased chondrocyte viability. Aberrant FSS induces remarkable inflammatory mediators production, leading to cartilage degeneration and degradation. In depth mechanistic study showed that FSS results in more than 10-fold upregulation of H3K4me3, and the modulatory effect of H3K4me3 on cartilage was obtained by directly targeting ZBTB20. Furthermore, Wnt signaling was strongly activated in high FSS-induced OA pathogenesis, and the negative impact of ZBTB20 on chondrocytes was also achieved through activating Wnt signaling pathway. Moreover, pharmacological inhibition of H3K4me3 activation by MM-102 or treatment with Wnt pathway inhibitor LF3 could effectively alleviate the destructive effect of FSS on chondrocytes. In vivo UAC mice model validated the dysregulation of H3K4me3 and ZBTB20 in aberrant force-induced cartilage pathogenesis. Conclusion Through the combination of in vitro FSS model and in vivo UAC model, KMT2B-H3K4me3-ZBTB20 axis was first identified in aberrant FSS-induced cartilage pathogenesis, which may provide evidences for epigenetic-based therapy in the future.