title: Identification of host miRNAs that may limit human rhinovirus replication.
abstract: OBJECTIVE
  To test whether the replication of human rhinovirus (HRV) is regulated by microRNAs in human bronchial epithelial cells.
  
  
  METHODS
  For the present study, the human cell line BEAS-2B (derived from normal human bronchial epithelial cells) was adopted. DICER knock-down, by siRNA transfection in BEAS-2B cells, was performed in order to inhibit microRNA maturation globally. Alternatively, antisense oligonucleotides (anti-miRs) were transfected to inhibit the activity of specific microRNAs. Cells were infected with HRV-1B. Viral replication was assessed by measuring the genomic viral RNA by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Association between microRNA-induced-silencing-complex and viral RNA was detected by Ago2 co-immunoprecipitation followed by RT-qPCR. Targetscan v.6 was used to predict microRNA target sites on several HRV strains.
  
  
  RESULTS
  Here, we show that microRNAs affect replication of HRV-1B. DICER knock-down significantly reduced the expression of mature microRNAs in a bronchial epithelial cell line (BEAS-2B) and in turn, increased the synthesis of HRV-1B RNA. Additionally, HRV-1B RNA co-immunoprecipitated with argonaute 2 protein, an important effector for microRNA activity suggesting that microRNAs bind to viral RNA during infection. In order to identify specific microRNAs involved in this interaction, we employed bioinformatics analysis, and selected a group of microRNAs that have been reported to be under-expressed in asthmatic bronchial epithelial cells and were predicted to target different strains of rhinoviruses (HRV-1B, -16, -14, -27). Our results suggest that, out of this group of microRNAs, miR-128 and miR-155 contribute to the innate defense against HRV-1B: transfection of specific anti-miRs increased viral replication, as anticipated in-silico.
  
  
  CONCLUSIONS
  Taken together, our results suggest that pathological changes in microRNA expression, as already reported for asthma or chronic obstructive pulmonary disease have the potential to affect Rhinovirus replication and therefore may play a role in virus-induced exacerbations.
authors.affiliation: Victor Paky Bondanese, LGL, LabEx LIO, ENS Lyon, Site Monod, 69364 Lyon, France.
authors.forename: Victor Paky
authors.initials: VP
authors.lastname: Bondanese
authors.affiliation: Victor Paky Bondanese, LGL, LabEx LIO, ENS Lyon, Site Monod, 69364 Lyon, France.
authors.forename: Ana
authors.initials: A
authors.lastname: Francisco-Garcia
authors.affiliation: Victor Paky Bondanese, LGL, LabEx LIO, ENS Lyon, Site Monod, 69364 Lyon, France.
authors.forename: Nicole
authors.initials: N
authors.lastname: Bedke
authors.affiliation: Victor Paky Bondanese, LGL, LabEx LIO, ENS Lyon, Site Monod, 69364 Lyon, France.
authors.forename: Donna E
authors.initials: DE
authors.lastname: Davies
authors.affiliation: Victor Paky Bondanese, LGL, LabEx LIO, ENS Lyon, Site Monod, 69364 Lyon, France.
authors.forename: Tilman
authors.initials: T
authors.lastname: Sanchez-Elsner
keywords: Antiviral innate immunity; Human Rhinovirus; Lentiviral transduction; MiR-155; MicroRNAs; SiRNA