miR-483 Targeting of CTGF Suppresses Endothelial-to-Mesenchymal Transition: Therapeutic Implications in Kawasaki Disease.
miR-483 Targeting of CTGF Suppresses Endothelial-to-Mesenchymal Transition: Therapeutic Implications in Kawasaki Disease.
复制标题
miR-483 靶向 CTGF 抑制内皮向间质转化：川崎病的治疗意义
DOI：
10.1161/circresaha.116.310233
复制
发表时间：
2017-01-20
影响因子：
20.1
通讯作者：
Shyy JY
中科院分区：
文献类型：
--
作者：
He M;Chen Z;Martin M;Zhang J;Sangwung P;Woo B;Tremoulet AH;Shimizu C;Jain MK;Burns JC;Shyy JY
内皮细胞向间充质转化（EndoMT）与急性川崎病（KD）患者中肌成纤维细胞样细胞介导的冠状动脉壁损伤有关，KD尸检组织中结缔组织生长因子（CTGF）和EndoMT标记物染色阳性即可证明这一点。然而，关于EndoMT参与KD的分子基础知之甚少。我们研究了微小RNA（miRNA）对CTGF的调节以及随后的KD发病机制中的EndoMT。同时，研究了他汀类药物治疗对这一过程的调节。将来自健康儿童和KD受试者的血清与人脐静脉内皮细胞（HUVECs）孵育。使用RT-qPCR、ELISA和蛋白质印迹定量心血管疾病相关的miRNA、CTGF和EndoMT标志物。与健康对照相比，与来自急性KD患者的血清孵育的HUVEC具有减少的miR-483、增加的CTGF和增加的EndoMT标记物。生物信息学分析和功能验证表明，Krüppel样因子4（KLF 4）可反式激活miR-483，进而靶向CTGF mRNA的3′非翻译区。KLF 4或pre-miR-483的过表达抑制了体外和体内EC中CTGF的表达，而KLF 4或anti-miR-483的敲低则增强了CTGF的表达。此外，阿托伐他汀，目前正在KD儿童的I/IIa期临床试验中进行测试，诱导KLF 4-miR-483，其抑制EC中的CTGF和EndoMT。KD血清抑制EC中的KLF 4-miR-483轴，导致CTGF表达增加和EndoMT诱导。内皮细胞的这种有害过程可能导致KD患者的冠状动脉异常。他汀类药物治疗可使急性KD患者受益，部分原因是通过恢复KLF 4-miR-483表达。NCT01431105
Endothelial-to-mesenchymal transition (EndoMT) is implicated in myofibroblast-like cell-mediated damage to the coronary arterial wall in acute Kawasaki disease (KD) patients, as evidenced by positive staining for connective tissue growth factor (CTGF) and EndoMT markers in KD autopsy tissues. However, little is known about the molecular basis of EndoMT involved in KD. We investigated the microRNA (miRNA) regulation of CTGF and the consequent EndoMT in KD pathogenesis. As well, the modulation of this process by statin therapy was studied. Sera from healthy children and KD subjects were incubated with human umbilical vein endothelial cells (HUVECs). Cardiovascular disease-related miRNAs, CTGF, and EndoMT markers were quantified using RT-qPCR, ELISA, and Western blotting. Compared to healthy controls, HUVEC incubated with sera from acute KD patients had decreased miR-483, increased CTGF, and increased EndoMT markers. Bioinformatics analysis followed by functional validation demonstrated that Krüppel-like factor 4 (KLF4) transactivates miR-483, which in turn targets the 3′ untranslated region of CTGF mRNA. Overexpression of KLF4 or pre-miR-483 suppressed, whereas knockdown of KLF4 or anti-miR-483 enhanced, CTGF expression in ECs in vitro and in vivo. Furthermore, atorvastatin, currently being tested in a Phase I/IIa clinical trial in KD children, induced KLF4-miR-483, which suppressed CTGF and EndoMT in ECs. KD sera suppress the KLF4-miR-483 axis in ECs leading to increased expression of CTGF and induction of EndoMT. This detrimental process in the endothelium may contribute to coronary artery abnormalities in KD patients. Statin therapy may benefit acute KD patients, in part through the restoration of KLF4-miR-483 expression. NCT01431105