Transforming Growth Factor-β1 Selectively Recruits microRNAs to the RNA-Induced Silencing Complex and Degrades CFTR mRNA under Permissive Conditions in Human Bronchial Epithelial Cells
Transforming Growth Factor-β1 Selectively Recruits microRNAs to the RNA-Induced Silencing Complex and Degrades CFTR mRNA under Permissive Conditions in Human Bronchial Epithelial Cells
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DOI：
10.3390/ijms20194933
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发表时间：
2019-10-01
影响因子：
5.6
通讯作者：
Swiatecka-Urban, Agnieszka
中科院分区：
文献类型：
--
作者：
Mitash, Nilay;Mu, Fangping;Swiatecka-Urban, Agnieszka
囊性纤维化跨膜传导调节因子（CFTR）基因突变导致囊性纤维化（CF）。最常见的突变F508 del抑制CFTR蛋白的折叠和加工。FDA批准的校正剂挽救F508 del-CFTR蛋白的生物合成加工，而增效剂改善挽救的CFTR通道功能。在许多CF患者中过度表达的转化生长因子（TGF-β 1）通过抑制CFTR mRNA在体外阻断校正剂/增强剂的拯救。增加的TGF-β 1信号传导和获得性CFTR功能障碍存在于其他肺部疾病中。为了研究TGF-β 1抑制CFTR的机制，我们在来自50多个供体的原代人支气管上皮细胞中使用了分子、生物化学和功能方法。TGF-β 1使慢性疾病（包括CF）肺细胞中的CFTR mRNA不稳定，新一代校正剂对F508 del-CFTR的拯救受损。TGF-β 1增加了经验证为CFTR抑制剂的选定微（mi）RNA的活性库，将它们招募到RNA诱导的沉默复合物（RISC）中。F508 del-CFTR的表达全面调节TGF-β 1诱导的miRNA景观变化，创造了F508 del-CFTR mRNA降解所需的宽松环境。总之，TGF-β 1可能会阻碍CF患者的校正剂/增强剂治疗的全部益处。研究疾病特异性条件下RISC的miRNA募集可能有助于更好地表征TGF-β 1用于破坏CFTR mRNA稳定性的miRNA。
Mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene lead to cystic fibrosis (CF). The most common mutation F508del inhibits folding and processing of CFTR protein. FDA-approved correctors rescue the biosynthetic processing of F508del-CFTR protein, while potentiators improve the rescued CFTR channel function. Transforming growth factor (TGF-beta 1), overexpressed in many CF patients, blocks corrector/potentiator rescue by inhibiting CFTR mRNA in vitro. Increased TGF-beta 1 signaling and acquired CFTR dysfunction are present in other lung diseases. To study the mechanism of TGF-beta 1 repression of CFTR, we used molecular, biochemical, and functional approaches in primary human bronchial epithelial cells from over 50 donors. TGF-beta 1 destabilized CFTR mRNA in cells from lungs with chronic disease, including CF, and impaired F508del-CFTR rescue by new-generation correctors. TGF-beta 1 increased the active pool of selected micro(mi)RNAs validated as CFTR inhibitors, recruiting them to the RNA-induced silencing complex (RISC). Expression of F508del-CFTR globally modulated TGF-beta 1-induced changes in the miRNA landscape, creating a permissive environment required for degradation of F508del-CFTR mRNA. In conclusion, TGF-beta 1 may impede the full benefit of corrector/potentiator therapy in CF patients. Studying miRNA recruitment to RISC under disease-specific conditions may help to better characterize the miRNAs utilized by TGF-beta 1 to destabilize CFTR mRNA.