MicroRNA-221 modulates RSV replication in human bronchial epithelium by targeting NGF expression.
MicroRNA-221 modulates RSV replication in human bronchial epithelium by targeting NGF expression.
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DOI：
10.1371/journal.pone.0030030
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Piedimonte G
中科院分区：
文献类型：
--
作者：
Othumpangat S;Walton C;Piedimonte G
生命早期呼吸道合胞病毒 (RSV) 感染与人类支气管上皮中原型神经营养蛋白神经生长因子 (NGF) 及其同源受体的异常表达有关。然而，导致这一结果的一系列事件及其对病毒感染进展的功能影响尚未阐明。本研究试图检验以下假设：RSV 感染通过沉默特定 microRNA (miRNA) 的表达来调节人类呼吸道中的神经营养通路，并且这种效应通过干扰受感染细胞的程序性死亡来促进病毒生长。使用多重 qPCR 阵列筛选感染绿色荧光蛋白表达 RSV (rgRSV) 的人支气管上皮细胞，并分析受病毒显着影响的 miRNA 与编码神经营养因子或受体的 mRNA 的同源性。将选定的 miRNA 的模拟序列转染到未感染的支气管细胞中，以确认它们各自在基因和蛋白质水平上调节神经营养素表达的作用，并研究它们对细胞周期和病毒复制的影响。 RSV 引起 24 种 miRNA 下调和 2 种 miRNA 上调（p<0.01）。微阵列数据的同源性分析显示，其中 6 个 miRNA 表现出与 NGF 和/或其同源受体 TrKA 和 p75NTR 之一的高度互补性。在所选的miRNA中，miR-221被RSV显着下调，其在支气管上皮细胞中的转染最大限度地抑制了NGF和TrKA的基因和蛋白表达，增加了细胞凋亡，并降低了病毒复制和感染性。我们的数据表明，RSV 通过沉默 miR-221 表达来上调人类气道中的 NGF-TrKA 轴，这通过干扰受感染细胞的凋亡死亡来促进病毒复制。因此，将外源 miRNA 靶向递送至气道可能为未来基于 RNA 干扰的抗病毒治疗提供新策略。
Early-life infection by respiratory syncytial virus (RSV) is associated with aberrant expression of the prototypical neurotrophin nerve growth factor (NGF) and its cognate receptors in human bronchial epithelium. However, the chain of events leading to this outcome, and its functional implications for the progression of the viral infection, has not been elucidated. This study sought to test the hypothesis that RSV infection modulates neurotrophic pathways in human airways by silencing the expression of specific microRNAs (miRNAs), and that this effect favors viral growth by interfering with programmed death of infected cells. Human bronchial epithelial cells infected with green fluorescent protein-expressing RSV (rgRSV) were screened with multiplex qPCR arrays, and miRNAs significantly affected by the virus were analyzed for homology with mRNAs encoding neurotrophic factors or receptors. Mimic sequences of selected miRNAs were transfected into non-infected bronchial cells to confirm the role of each of them in regulating neurotrophins expression at the gene and protein level, and to study their influence on cell cycle and viral replication. RSV caused downregulation of 24 miRNAs and upregulation of 2 (p<0.01). Homology analysis of microarray data revealed that 6 of those miRNAs exhibited a high degree of complementarity to NGF and/or one of its cognate receptors TrKA and p75NTR. Among the selected miRNAs, miR-221 was significantly downregulated by RSV and its transfection in bronchial epithelial cells maximally inhibited gene and protein expression of NGF and TrKA, increased apoptotic cell death, and reduced viral replication and infectivity. Our data suggest that RSV upregulates the NGF-TrKA axis in human airways by silencing miR-221 expression, and this favors viral replication by interfering with the apoptotic death of infected cells. Consequently, the targeted delivery of exogenous miRNAs to the airways may provide a new strategy for future antiviral therapies based on RNA interference.
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