Overexpression of ubiquitin-specific peptidase 15 in systemic sclerosis fibroblasts increases response to transforming growth factor
Overexpression of ubiquitin-specific peptidase 15 in systemic sclerosis fibroblasts increases response to transforming growth factor
复制标题
DOI：
10.1093/rheumatology/key401
复制
发表时间：
2019-04-01
期刊：
影响因子：
5.5
通讯作者：
Lauwerys, Bernard R.
中科院分区：
文献类型：
--
作者：
Galant, Christine;Marchandise, Joel;Lauwerys, Bernard R.
目的 蛋白质的泛素化导致其被蛋白酶体降解，并受到泛素连接酶和底物特异性泛素特异性肽酶 (USP) 的调节。泛素化过程在细胞代谢和细胞周期的调节中也发挥着重要作用。在这里，我们发现几种 USP 的表达在 SSc 腱滑膜和皮肤活检中增加，并且我们证明 USP 抑制会减少原代成纤维细胞系中的 TGF-信号传导。方法使用从 SSc 腱滑膜样品中获得的总 RNA 进行高密度转录组研究。对腱滑膜和皮肤样本进行了验证性免疫染色实验。进行体外实验是为了研究 USP 调节对 TGF-刺激反应的影响。结果 SSc 患者的腱滑膜活检过度表达已知的疾病相关基因途径：纤维化、细胞因子和趋化因子、Wnt/TGF-信号传导，还有几种 USP。免疫组织化学实验证实在相同样本和 SSc 皮肤活检中检测到了 USP。将原代成纤维细胞系暴露于 TGF 诱导的 USP 基因表达。使用泛 USP 抑制剂可降低 TGF 刺激的成纤维细胞中 SMAD3 磷酸化以及 COL1A1、COL3A1 和纤连蛋白基因的表达。 USP抑制剂的作用导致SMAD3泛素化增加，并被蛋白酶体抑制剂阻断，从而证实了其作用的特异性。 结论 包括USP15在内的几种USP的过表达可放大TGF-β诱导的纤维化反应，是SSc的潜在治疗靶点。
Objective Ubiquitination of proteins leads to their degradation by the proteasome, and is regulated by ubiquitin ligases and substrate-specific ubiquitin-specific peptidases (USPs). The ubiquitination process also plays important roles in the regulation of cell metabolism and cell cycle. Here, we found that the expression of several USPs is increased in SSc tenosynovial and skin biopsies, and we demonstrated that USP inhibition decreases TGF- signalling in primary fibroblast cell lines.Methods High-density transcriptomic studies were performed using total RNA obtained from SSc tenosynovial samples. Confirmatory immunostaining experiments were performed on tenosynovial and skin samples. In vitro experiments were conducted in order to study the influence of USP modulation on responses to TGF- stimulation.Results Tenosynovial biopsies from SSc patients overexpressed known disease-associated gene pathways: fibrosis, cytokines and chemokines, and Wnt/TGF- signalling, but also several USPs. Immunohistochemistry experiments confirmed the detection of USPs in the same samples, and in SSc skin biopsies. Exposure of primary fibroblast cell lines to TGF- induced USP gene expression. The use of a pan-USP inhibitor decreased SMAD3 phosphorylation, and expression of COL1A1, COL3A1 and fibronectin gene expression in TGF--stimulated fibroblasts. The effect of the USP inhibitor resulted in increased SMAD3 ubiquitination, and was blocked by a proteasome inhibitor, thereby confirming the specificity of its action.Conclusion Overexpression of several USPs, including USP15, amplifies fibrotic responses induced by TGF-, and is a potential therapeutic target in SSc.