Shi-Wen Xu, Racanelli Michael, Ali Aaisham, Simon Amara, Quesnel Katherine, Stratton Richard J, Leask Andrew
UCL Division of Medicine, Centre for Rheumatology, University College London, Royal Free Campus, Rowland Hill St, London, NW3 2PF, UK.
Department of Physiology and Pharmacology, University of Western Ontario, London, ON, N6A 5C1, Canada.
J Cell Commun Signal. 2021 Mar;15(1):71-80. doi: 10.1007/s12079-020-00596-x. Epub 2021 Jan 4.
Fibrosis is perpetuated by an autocrine, pro-adhesive signaling loop maintained by the synthetic and contractile abilities of myofibroblasts and the stiff, highly-crosslinked extracellular matrix. Transcriptional complexes that are exquisitely responsive to mechanotransduction include the co-activator YAP1, which regulates the expression of members of the CCN family of matricellular proteins such as CCN2 and CCN1. Although selective YAP1 inhibitors exist, the effect of these inhibitors on profibrotic gene expression in fibroblasts is largely unknown, and is the subject of our current study. Herein, we use genome-wide expression profiling, real-time polymerase chain reaction and Western blot analyses, cell migration and collagen gel contraction assays to assess the ability of a selective YAP inhibitor verteporfin (VP) to block fibrogenic activities in dermal fibroblasts from healthy individual human controls and those from isolated from fibrotic lesions of patients with diffuse cutaneous systemic sclerosis (dcSSc). In control fibroblasts, VP selectively reduced expression of fibrogenic genes and also blocked the ability of TGFbeta to induce actin stress fibers in dermal fibroblasts. VP also reduced the persistent profibrotic phenotype of dermal fibroblasts cultured from fibrotic lesions of patients with dcSSc. Our results are consistent with the notion that, in the future, YAP1 inhibitors may represent a novel, valuable method of treating fibrosis as seen in dcSSc.
纤维化通过一个自分泌的、促进黏附的信号回路持续存在，该回路由肌成纤维细胞的合成和收缩能力以及僵硬、高度交联的细胞外基质维持。对机械转导高度敏感的转录复合物包括共激活因子YAP1，它调节基质细胞蛋白CCN家族成员（如CCN2和CCN1）的表达。尽管存在选择性YAP1抑制剂，但这些抑制剂对成纤维细胞中促纤维化基因表达的影响在很大程度上尚不清楚，这也是我们当前研究的主题。在此，我们使用全基因组表达谱分析、实时聚合酶链反应和蛋白质印迹分析、细胞迁移和胶原凝胶收缩试验，来评估选择性YAP抑制剂维替泊芬（VP）阻断来自健康个体对照的皮肤成纤维细胞以及来自弥漫性皮肤系统性硬化症（dcSSc）患者纤维化病变处分离的皮肤成纤维细胞的纤维化活性的能力。在对照成纤维细胞中，VP选择性降低促纤维化基因的表达，还阻断了TGFβ诱导皮肤成纤维细胞中肌动蛋白应力纤维的能力。VP还降低了从dcSSc患者纤维化病变处培养的皮肤成纤维细胞持续的促纤维化表型。我们的结果与以下观点一致，即未来YAP1抑制剂可能代表一种治疗dcSSc中所见纤维化的新的、有价值的方法。
