Involvement of YTHDF1 in renal fibrosis progression via up‐regulating YAP
Involvement of YTHDF1 in renal fibrosis progression via up‐regulating YAP
复制标题
DOI：
10.1096/fj.202100172rr
复制
发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Jia Xing;Yuping He;Kai-Yue Wang;Peng Wan;Xiaoyue Zhai
中科院分区：
文献类型：
--
作者：
Jia Xing;Yuping He;Kai-Yue Wang;Peng Wan;Xiaoyue Zhai
肾纤维化是一种进行性、致命性的肾脏疾病，其特征是肌成纤维细胞在肾间质和肾小球中异常积聚，产生过量的细胞外基质（ECM）。Yes相关蛋白（YAP）被认为是肌成纤维细胞转化的关键调节剂，但其上游调节剂仍是一个谜。本研究通过生物信息学分析探讨了m6A甲基化在肾纤维化过程中的作用，我们发现m6A甲基化调节剂YTHDF1是肾纤维化的关键因素，因为它在人类纤维化肾脏中高度表达，并且与YAP有显著的校正作用。免疫荧光还显示了它们在人纤维化肾脏中的共定位。然后，我们发现YTHDF1在单侧输尿管梗阻（UUO）、高剂量叶酸或单侧缺血再灌注损伤诱导的纤维化小鼠肾脏中也上调，进一步支持了YTHDF1在肾纤维化中的因果作用。与这一观点一致的是，YTHDF1敲低减轻了转化生长因子- β诱导的培养细胞和UUO小鼠模型中肾纤维化的进展。同时，YTHDF1被抑制时，YAP也相应下调。此外，通过RNA binding Protein Immunoprecipitation检测YTHDF1与YAP mRNA的特异性结合，YTHDF1过表达质粒诱导的纤维化相关分子在培养细胞中的上调被YAP siRNA减弱。综上所述，我们的数据强调了YTHDF1作为肾纤维化指标的潜在效用，并表明抑制YTHDF1可能是通过下调YAP来缓解肾纤维化的一种有前景的治疗策略。
Renal fibrosis is a progressive, fatal renal disease characterized by the aberrant accumulation of myofibroblasts that produce excess extracellular matrix (ECM) in the renal interstitium and glomeruli. Yes‐associated protein (YAP) has been regarded as a crucial modulator in myofibroblast transformation, but its upstream regulator remains a mystery. In the present study investigating the participation of m6A methylation during renal fibrosis through bioinformatics analysis, we identified YTHDF1, a modulator of m6A methylation, as a key contributor for renal fibrosis because it was highly expressed in human fibrotic kidneys and had a significant correction with YAP. Their co‐localization in human fibrotic kidneys was additionally shown by immunofluorescence. We then found that YTHDF1 was also up‐regulated in fibrotic mouse kidneys induced by unilateral ureteral obstruction (UUO), high‐dose folic acid administration, or the unilateral ischemia‐reperfusion injury, further supporting a causal role of YTHDF1 during renal fibrosis. Consistent with this notion, YTHDF1 knockdown alleviated the progression of renal fibrosis both in cultured cells induced by transforming growth factor‐beta administration and in the UUO mouse model. Meanwhile, YAP was accordingly down‐regulated when YTHDF1 was inhibited. Furthermore, the specific binding of YTHDF1 to YAP mRNA was detected using RNA Binding Protein Immunoprecipitation, and the up‐regulation of fibrotic related molecules in cultured cells induced by YTHDF1 over‐expression plasmid was attenuated by YAP siRNA. Taken together, our data highlight the potential utility of YTHDF1 as an indicator for renal fibrosis and suggest that YTHDF1 inhibition might be a promising therapeutic strategy to alleviate renal fibrosis via downregulating YAP.