Role of Substratum Stiffness in Modulating Genes Associated with Extracellular Matrix and Mechanotransducers YAP and TAZ
Role of Substratum Stiffness in Modulating Genes Associated with Extracellular Matrix and Mechanotransducers YAP and TAZ
复制标题
DOI：
10.1167/iovs.12-11007
复制
发表时间：
2013-01-01
影响因子：
4.4
通讯作者：
Russell, Paul
中科院分区：
文献类型：
--
作者：
Raghunathan, Vijay Krishna;Morgan, Joshua T.;Russell, Paul
目的.原发性开角型青光眼的特征在于增加的对房水流出的阻力和更硬的人小梁网（HTM）。Yorkie家族的两个同源基因是Yes相关蛋白（雅普）和转录辅激活因子（TAZ），它们是细胞外微环境的机械转导子和转录辅激活因子。在这里，我们探索基质硬度如何调节HTM细胞中的雅普/TAZ途径和细胞外基质基因，以及这如何在青光眼的发生和进展中发挥作用。方法。将来自正常供体的HTM细胞在模拟正常（5 kPa）和昏迷（75 kPa）HTM的硬度的水凝胶上培养。测定雅普/TAZ相关基因表达和激素反应性的变化。此外，在雅普沉默后测定转氨酶-2的表达。雅普和TAZ均在人小梁网细胞中表达。在体外，雅普和TAZ是负调控的基质刚度。雅普和14-3-3 sigma在较硬的底物上不同程度地下调; TAZ、组织转氨酶（TGM 2）和可溶性卷曲相关蛋白-1（sFRP-1）显著上调。CTGF的表达似乎被雅普和TAZ改变差异。肌球蛋白和血管生成素样7的表达在地塞米松的反应更显着更硬的基板。我们证明了当雅普被小干扰RNA沉默时，雅普对TGM 2的直接作用。雅普/TAZ和ECM相关基因的表达受生理相关底物的影响。雅普在较软基质上的细胞中上调。较硬的基质导致经典Wnt调节剂TAZ和sFRP-1的上调，因此可能影响青光眼的进展。这些结果证明了雅普/TAZ在HTM中的重要性，并表明它们在青光眼中的作用。（Invest Ophthalmol维斯科学。2013; 54：378-386）DOI：10.1167/iovs.12-11007
PURPOSE. Primary open-angle glaucoma is characterized by increased resistance to aqueous humor outflow and a stiffer human trabecular meshwork (HTM). Two Yorkie homologues, Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif, encoded by WWTR1 (TAZ), are mechanotransducers of the extracellular-microenvironment and coactivators of transcription. Here, we explore how substratum stiffness modulates the YAP/TAZ pathway and extracellular matrix genes in HTM cells and how this may be play a role in the onset and progression of glaucoma.METHODS. HTM cells from normal donors were cultured on hydrogels mimicking the stiffness of normal (5 kPa) and glaucomatous (75 kPa) HTM. Changes in expression of YAP/TAZ related genes and steroid responsiveness were determined. Additionally, transglutaminase-2 expression was determined after YAP silencing.RESULTS. YAP and TAZ are both expressed in human trabecular meshwork cells. In vitro, YAP and TAZ were inversely regulated by substratum stiffness. YAP and 14-3-3 sigma were downregulated to different extents on stiffer substrates; TAZ, tissue transglutaminase (TGM2), and soluble frizzled-related protein-1 (sFRP-1) were significantly upregulated. CTGF expression appeared to be altered differentially by both YAP and TAZ. Myocilin and angiopoietin-like 7 expression in response to dexamethasone was more pronounced on stiffer substrates. We demonstrated a direct effect by YAP on TGM2 when YAP was silenced by small interfering RNA.CONCLUSIONS. The expression of YAP/TAZ and ECM-related-genes is impacted on physiologically relevant substrates. YAP was upregulated in cells on softer substrates. Stiffer substrates resulted in upregulation of canonical Wnt modulators, TAZ and sFRP-1, and thus may influence the progression of glaucoma. These results demonstrate the importance of YAP/TAZ in the HTM and suggest their role in glaucoma. (Invest Ophthalmol Vis Sci. 2013; 54: 378-386) DOI: 10.1167/iovs.12-11007