MicroRNA-221 Modulates RSV Replication in Human Bronchial Epithelium by Targeting NGF Expression 英文参考文献.docVIP

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MicroRNA-221 Modulates RSV Replication in Human Bronchial Epithelium by Targeting NGF Expression 英文参考文献.doc

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MicroRNA-221 Modulates RSV Replication in Human Bronchial Epithelium by Targeting NGF Expression 英文参考文献

MicroRNA-221ModulatesRSVReplicationinHuman BronchialEpitheliumbyTargetingNGFExpression SreekumarOthumpangat,CherylWalton,GiovanniPiedimonte* DepartmentofPediatricsandPediatricResearchInstitute,WestVirginiaUniversitySchoolofMedicine,Morgantown,WestVirginia,UnitedStatesofAmerica Abstract Background:Early-lifeinfectionbyrespiratorysyncytialvirus(RSV)isassociatedwithaberrantexpressionoftheprototypical neurotrophinnervegrowthfactor(NGF)anditscognatereceptorsinhumanbronchialepithelium.However,thechainof events leading to this outcome, and its functional implications for the progression of the viral infection, has not been elucidated.ThisstudysoughttotestthehypothesisthatRSVinfectionmodulatesneurotrophicpathwaysinhumanairways by silencing the expression of specific microRNAs (miRNAs), and that this effect favors viral growth by interfering with programmeddeathofinfectedcells. Methodology: 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- infectedbronchialcellstoconfirmtheroleofeachoftheminregulatingneurotrophinsexpressionatthegeneandprotein level,andtostudytheirinfluenceoncellcycleandviralreplication. Principal Findings: 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 cognatereceptorsTrKAandp75NTR.AmongtheselectedmiRNAs,miR-221wassignificantlydownregulatedbyRSVandits transfection in bronchial epithelial cells maximally inhibited gene and protein expression of NGF and TrKA, increased apoptoticcelldeath,andreducedviralreplicationandinfectivity. Conclusions/Significance:OurdatasuggestthatRSVupregulatestheNGF-TrKAaxisinhumanairwaysbysilencingmi