title: Investigating the Influence of Ribavirin on Human Respiratory Syncytial Virus RNA Synthesis by Using a High-Resolution Transcriptome Sequencing Approach.
abstract: Human respiratory syncytial virus (HRSV) is a major cause of serious respiratory tract infection. Treatment options include administration of ribavirin, a purine analog, although the mechanism of its anti-HRSV activity is unknown. We used transcriptome sequencing (RNA-seq) to investigate the genome mutation frequency and viral mRNA accumulation in HRSV-infected cells that were left untreated or treated with ribavirin. In the absence of ribavirin, HRSV-specific transcripts accounted for up to one-third of total RNA reads from the infected-cell RNA population. Ribavirin treatment resulted in a >90% reduction in abundance of viral mRNA reads, while at the same time no such reduction was detected for the abundance of cellular transcripts. The presented data reveal that ribavirin significantly increases the frequency of HRSV-specific RNA mutations, suggesting a direct influence on the fidelity of the HRSV polymerase. The presented data show that transitions and transversions occur during HRSV replication and that these changes occur in hot spots along the HRSV genome. Examination of nucleotide substitution rates in the viral genome indicated an increase in the frequency of transition but not transversion mutations in the presence of ribavirin. In addition, our data indicate that in the continuous cell types used and at the time points analyzed, the abundances of some HRSV mRNAs do not reflect the order in which the mRNAs are transcribed.
  
  
  
  Human respiratory syncytial virus (HRSV) is a major pediatric pathogen. Ribavirin can be used in children who are extremely ill to reduce the amount of virus and to lower the burden of disease. Ribavirin is used as an experimental therapy with other viruses. The mechanism of action of ribavirin against HRSV is not well understood, although it is thought to increase the mutation rate of the viral polymerase during replication. To investigate this hypothesis, we used a high-resolution approach that allowed us to determine the genetic sequence of the virus to a great depth of coverage. We found that ribavirin did not cause a detectable change in the relative amounts of viral mRNA transcripts. However, we found that ribavirin treatment did indeed cause an increase in the number of mutations, which was associated with a decrease in virus production.
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom.|Health Protection Research Unit in Emerging and Zoonotic Infections, Liverpool, United Kingdom.
authors.forename: Waleed
authors.initials: W
authors.lastname: Aljabr
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Olivier
authors.initials: O
authors.lastname: Touzelet
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom.|Health Protection Research Unit in Emerging and Zoonotic Infections, Liverpool, United Kingdom.
authors.forename: Georgios
authors.initials: G
authors.lastname: Pollakis
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Weining
authors.initials: W
authors.lastname: Wu
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Diane C
authors.initials: DC
authors.lastname: Munday
authors.affiliation: Centre for Genomic Research, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Margaret
authors.initials: M
authors.lastname: Hughes
authors.affiliation: Centre for Genomic Research, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Christiane
authors.initials: C
authors.lastname: Hertz-Fowler
authors.affiliation: Centre for Genomic Research, University of Liverpool, Liverpool, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Kenny
authors.affiliation: School of Medicine, Boston University, Boston, Massachusetts, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Fearns
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom.
authors.forename: John N
authors.initials: JN
authors.lastname: Barr
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Bristol, United Kingdom d.a.matthews@bristol.ac.uk julian.hiscox@liverpool.ac.uk.
authors.forename: David A
authors.initials: DA
authors.lastname: Matthews
authors.affiliation: Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom d.a.matthews@bristol.ac.uk julian.hiscox@liverpool.ac.uk.|Health Protection Research Unit in Emerging and Zoonotic Infections, Liverpool, United Kingdom.
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D016679:Genome, Viral / Q000187:drug effects; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism; D009154:Mutation; D012367:RNA, Viral / Q000235:genetics*; D018357:Respiratory Syncytial Virus Infections / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D012254:Ribavirin / Q000494:pharmacology*; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D010948:Viral Plaque Assay; D053585:Virus Attachment / Q000187:drug effects; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral; D012254:Ribavirin; D016899:Interferon-beta