Frömmichen Janne, Bungert Emma, Ströble Jeanne, Gläser Moritz, Gottwald Charlotte, Zeqiri Kosovare, Reinhard Thomas, Lübke Jan, Schlunck Günther, Wiedenmann Cornelius Jakob
Eye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Invest Ophthalmol Vis Sci. 2025 Apr 1;66(4):17. doi: 10.1167/iovs.66.4.17.
Postoperative scarring remains the major challenge in achieving long-term success after glaucoma filtration surgery. In a previous study, we showed that slow continuous fluid flow is sufficient to induce fibrotic responses in human tenon fibroblasts (HTFs) in two-dimensional (2D) and three-dimensional (3D) in vitro models. In the present study, we investigated the role of the mechanosensitive Yes-associated protein (YAP) and transcriptional coactivator (TAZ) signaling pathway in flow-induced fibrosis.
HTFs were exposed to continuous fluid flow for 48 or 72 hours in the presence or absence of the YAP/TAZ-transcriptional enhanced associated domain inhibitor verteporfin (VP). In a 2D model, the F-actin cytoskeleton, fibronectin 1 (FN1), YAP, and TAZ were visualized by confocal immunofluorescence microscopy. In a 3D model, mRNA was extracted, and the expression of fibrosis-associated genes was detected by quantitative PCR.
HTFs exposed to slow fluid flow showed increased staining intensities for YAP/TAZ. Inhibition of YAP/TAZ by VP slightly reduced flow-induced fibrotic changes in the 2D model. The flow-induced increase in the expression of the extracellular matrix (ECM) genes COL1A1, CTGF, and FN1 was significantly inhibited by VP in the 3D model.
Slow interstitial fluid flow activates the YAP/TAZ pathway. VP exerts antifibrotic potential by reducing morphologic changes and suppressing the expression of ECM genes induced by flow. Therefore, YAP/TAZ inhibition may exhibit therapeutic potential after glaucoma filtration surgery by inhibiting fibrotic changes induced by mechanical stimuli.
术后瘢痕形成仍然是青光眼滤过手术取得长期成功的主要挑战。在先前的一项研究中，我们表明，在二维（2D）和三维（3D）体外模型中，缓慢持续的流体流动足以诱导人Tenon成纤维细胞（HTF）发生纤维化反应。在本研究中，我们调查了机械敏感性Yes相关蛋白（YAP）和转录共激活因子（TAZ）信号通路在流动诱导的纤维化中的作用。
在存在或不存在YAP/TAZ转录增强相关结构域抑制剂维替泊芬（VP）的情况下，将HTF暴露于持续流体流动48或72小时。在2D模型中，通过共聚焦免疫荧光显微镜观察F-肌动蛋白细胞骨架、纤连蛋白1（FN1）、YAP和TAZ。在3D模型中，提取mRNA，并通过定量PCR检测纤维化相关基因的表达。
暴露于缓慢流体流动的HTF显示YAP/TAZ的染色强度增加。在2D模型中，VP对YAP/TAZ的抑制略微减少了流动诱导的纤维化变化。在3D模型中，VP显著抑制了流动诱导的细胞外基质（ECM）基因COL1A1、CTGF和FN1表达的增加。
缓慢的间质液流动激活YAP/TAZ通路。VP通过减少形态学变化和抑制流动诱导的ECM基因表达发挥抗纤维化潜力。因此，抑制YAP/TAZ可能通过抑制机械刺激诱导的纤维化变化而在青光眼滤过手术后展现出治疗潜力。
