title: Targeting of protease 2A genome by single and multiple siRNAs as a strategy to impair CVB3 life cycle in permissive HeLa cells.
abstract: Dilated cardiomyopathy as a consequence of myocarditis induced by Coxsackieviruses B3 (CVB3) is a worldwide cause of morbidity and death for which current therapies have shown only modest effectiveness. In the present study we tested the efficacy of a novel CVB3 small interfering RNA (siRNA), directed against the protease 2A genome region (siRNA 3626), alone or in combination with another recently described protease 2A siRNA (siRNA 3541). siRNAs (400 nM) were delivered by lipofection to HeLa cells, which were then infected by CVB3 and analyzed. Under optimized transfection conditions, siRNA 3626 markedly improved the viability of CVB3-infected HeLa cells. This was due to a drastic reduction in the amounts of CVB3 genome, viral capsid protein VP1 and vital virions. Finally, the combination of siRNA 3626/3541 did not potentiate the anti-CVB3 effect compared with an equimolar concentration of either siRNA. In conclusion, these data show the potency of siRNA 3626 in downmodulating CVB3 infection together with the lack of any significant transfection-related toxicity. In addition to placing the selected siRNA 3626 among the most effective anti-CVB3 agents so far described, these data fully justify future attempts to further optimize the siRNA-mediated approach to CVB3-induced myocarditis.
authors.affiliation: Department of Molecular Pathology, Institute for Pathology, University Hospital of Tubingen, Germany.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Racchi
authors.forename: Karin
authors.initials: K
authors.lastname: Klingel
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kandolf
authors.forename: Gabriele
authors.initials: G
authors.lastname: Grassi
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D003384:Coxsackievirus Infections / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000235:genetics*; D029822:Enterovirus B, Human / Q000235:genetics*; D016679:Genome, Viral; D006367:HeLa Cells; D006801:Humans; D009205:Myocarditis / Q000235:genetics / Q000821:virology; D034741:RNA, Small Interfering / Q000494:pharmacology*; D014162:Transfection / Q000379:methods; D014764:Viral Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D034741:RNA, Small Interfering; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus