title: Small RNAs targeting the 5' end of the viral polymerase gene segments specifically interfere with influenza type A virus replication.
abstract: Human and avian influenza A viruses, associated with seasonal epidemics and occasionally with pandemics, have a high impact on public health. The development of new antivirals to counteract the emergence of drug resistant influenza virus variants is a main concern. The aim of this study was to develop systems for the efficient and stable expression of small therapeutic RNAs into influenza virus infected cells in order to get further insights on the efficacy of nucleic acid-based antiviral strategies. To this end, lentiviral vectors expressing either microRNAs or antisense-RNAs targeting the 5' end of the PA, PB1 and PB2 influenza virus genomic sequences were generated. Derivative recombinant lentiviral particles were employed to transduce the influenza virus highly susceptible human alveolar basal epithelial A549 cells. The expression of both RNA molecules led to a reduction up to 3 logs of the viral titer when transduced A549 cells were challenged with different human and avian subtypes of influenza type A virus. Importantly, no inhibition of influenza type B virus was observed. Overall our data support the development of nucleic acid-based antiviral strategies to control human and avian influenza A virus infection.
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Martelli
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Cristiano
authors.initials: C
authors.lastname: Salata
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Calistri
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Parolin
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
authors.forename: Alberta
authors.initials: A
authors.lastname: Azzi
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palù
authors.affiliation: Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy. Electronic address: simone.giannecchini@unifi.it.
authors.forename: Simone
authors.initials: S
authors.lastname: Giannecchini
mesh_terms: D020121:5' Untranslated Regions / Q000187:drug effects; D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D005822:Genetic Vectors / Q000494:pharmacology; D006801:Humans; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D016086:Lentivirus / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors* / Q000235:genetics; D000067492:RNAi Therapeutics; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D014779:Virus Replication
chemical_list: D020121:5' Untranslated Regions; D000998:Antiviral Agents; C061290:PA protein, influenza viruses; C057027:PB2 protein, Influenzavirus A; D016372:RNA, Antisense; D014764:Viral Proteins; C077346:influenza virus polymerase basic protein 1; D012324:RNA-Dependent RNA Polymerase
keywords: Antisense RNA; Influenza virus; Lentiviral vector; Packaging regions; Virus inhibition; microRNA