YAP activation inhibits inflammatory signalling and cartilage breakdown associated with reduced primary cilia expression.
YAP activation inhibits inflammatory signalling and cartilage breakdown associated with reduced primary cilia expression.
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DOI：
10.1016/j.joca.2022.11.001
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发表时间：
2022-11
影响因子：
7
通讯作者：
H. Meng;S. Fu;Marta B. Ferreira;Yu Hou;O. Pearce;Nuria Gavara;M. Knight
中科院分区：
文献类型：
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作者：
H. Meng;S. Fu;Marta B. Ferreira;Yu Hou;O. Pearce;Nuria Gavara;M. Knight
目的探讨YAP在调节软骨炎症和降解中的作用以及初级纤毛和相关的鞭毛内转运的参与。方法将分离的原代软骨细胞培养在不同硬度(6~1000kPa)的基质上，或用YAP激动剂溶血磷脂酸(LPA)或YAP拮抗剂维替普芬(VP)处理，或用YAP siRNA修饰，均为±1IL1β。测定一氧化氮(NO)和前列腺素E_2(PGE_2)释放以监测IL-1β反应。用YAP核浆比和YAP阳性细胞百分率来量化YAP活性。测试软骨移植的力学性能，以确认软骨的降解。结果LPA处理或增加聚二甲基硅氧烷底物硬度，激活YAP核的表达，并抑制IL1β诱导的NO和PGE2的释放。LPA治疗还可抑制IL-1β介导的软骨移植体内的炎症信号，防止基质降解和软骨生物力学损失。YAP的激活减少了初级纤毛的表达，在没有功能纤毛的情况下，YAP的敲除未能诱导炎症反应。结论YAP的药物激活和机械激活都能阻断IL1β诱导的促炎信号，防止软骨破坏和生物力学功能的丧失。这与初级纤毛表达减少有关，揭示了一种潜在的抗炎机制，具有治疗骨关节炎(OA)的新靶点。
ObjectiveTo clarify the role of YAP in modulating cartilage inflammation and degradation and the involvement of primary cilia and associated intraflagellar transport (IFT).MethodsIsolated primary chondrocytes were cultured on substrates of different stiffness (6–1000 kPa) or treated with YAP agonist lysophosphatidic acid (LPA) or YAP antagonist verteporfin (VP), or genetically modified by YAP siRNA, all ± IL1β. Nitric oxide (NO) and prostaglandin E2(PGE2) release were measured to monitor IL1β response. YAP activity was quantified by YAP nuclear/cytoplasmic ratio and percentage of YAP-positive cells. Mechanical properties of cartilage explants were tested to confirm cartilage degradation. The involvement of primary cilia and IFT was analysed using IFT88 siRNA and ORPK cells with hypomorphic mutation of IFT88.ResultsTreatment with LPA, or increasing polydimethylsiloxane (PDMS) substrate stiffness, activated YAP nuclear expression and inhibited IL1β-induced release of NO and PGE2, in isolated chondrocytes. Treatment with LPA also inhibited IL1β-mediated inflammatory signalling in cartilage explants and prevented matrix degradation and the loss of cartilage biomechanics. YAP activation reduced expression of primary cilia, knockdown of YAP in the absence of functional cilia/IFT failed to induce an inflammatory response.ConclusionsWe demonstrate that both pharmaceutical and mechanical activation of YAP blocks pro-inflammatory signalling induced by IL1β and prevents cartilage breakdown and the loss of biomechanical functionality. This is associated with reduced expression of primary cilia revealing a potential anti-inflammatory mechanism with novel therapeutic targets for treatment of osteoarthritis (OA).