tetano
Editor, Senior Moderator
J Virol. 2018 Nov 21. pii: JVI.01916-18. doi: 10.1128/JVI.01916-18. [Epub ahead of print]
[h=1]Efficient inhibition of avian and seasonal influenza A viruses by a virus-specific Dicer-substrate siRNA swarm in human monocyte-derived macrophages and dendritic cells.[/h] Jiang M[SUP]1,[/SUP][SUP]2[/SUP], ?sterlund P[SUP]3[/SUP], Westenius V[SUP]3[/SUP], Guo D[SUP]4[/SUP], Poranen MM[SUP]2[/SUP], Bamford DH[SUP]2[/SUP], Julkunen I[SUP]5[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza A viruses (IAVs) are viral pathogens that cause epidemics and occasional pandemics of significant mortality. The generation of efficacious vaccines and antiviral drugs remains a challenge due to the rapid appearance of new influenza types and antigenic variants. Consequently, novel strategies for the prevention and treatment of IAV infections are needed given the limitations of the presently available antivirals. Here, we used enzymatically produced IAV-specific double-stranded RNA (dsRNA) molecules and Giardia intestinalis Dicer for the generation of a swarm of small interfering RNA (siRNA) molecules. The siRNAs target multiple conserved genomic regions of the IAVs. In mammalian cells, the produced 25-27 nucleotide long siRNA molecules are processed by endogenous Dicer into 21-nucleotide siRNAs and are thus designated as Dicer-substrate siRNAs (DsiRNAs). We evaluated the efficacy of the above DsiRNA swarm at preventing IAV infections in human primary monocyte-derived macrophages and dendritic cells. The replication of different IAV strains, including avian influenza H5N1 and H7N9 viruses, was significantly inhibited by pre-transfection of the cells with the IAV-specific DsiRNA swarm. Up to 7 orders of magnitude inhibition of viral RNA expression was observed, which led to a dramatic inhibition of IAV protein synthesis and virus production. The IAV-specific DsiRNA swarm inhibited virus replication directly through the RNA interference pathway although a weak induction of innate interferon responses was detected. Our results provide direct evidence for the feasibility of the siRNA strategy and the potency of DsiRNA swarms in the prevention and treatment of influenza, including the highly pathogenic avian influenza viruses.IMPORTANCE In spite of the enormous amount of research, influenza virus is still one of the major challenges for medical virology due to its capacity to generate new variants, which potentially lead to severe epidemics and pandemics. We demonstrated here that a swarm of small interfering RNA (siRNA) molecules, including more than one hundred different antiviral RNA molecules targeting the most conserved regions of influenza A virus genome, could efficiently inhibit the replication of all tested avian and seasonal influenza A variants in human primary monocyte-derived macrophages and dendritic cells. The wide antiviral spectrum makes the virus-specific siRNA swarm a potentially efficient treatment modality against both avian and seasonal influenza viruses.
PMID: 30463970 DOI: 10.1128/JVI.01916-18
[h=1]Efficient inhibition of avian and seasonal influenza A viruses by a virus-specific Dicer-substrate siRNA swarm in human monocyte-derived macrophages and dendritic cells.[/h] Jiang M[SUP]1,[/SUP][SUP]2[/SUP], ?sterlund P[SUP]3[/SUP], Westenius V[SUP]3[/SUP], Guo D[SUP]4[/SUP], Poranen MM[SUP]2[/SUP], Bamford DH[SUP]2[/SUP], Julkunen I[SUP]5[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza A viruses (IAVs) are viral pathogens that cause epidemics and occasional pandemics of significant mortality. The generation of efficacious vaccines and antiviral drugs remains a challenge due to the rapid appearance of new influenza types and antigenic variants. Consequently, novel strategies for the prevention and treatment of IAV infections are needed given the limitations of the presently available antivirals. Here, we used enzymatically produced IAV-specific double-stranded RNA (dsRNA) molecules and Giardia intestinalis Dicer for the generation of a swarm of small interfering RNA (siRNA) molecules. The siRNAs target multiple conserved genomic regions of the IAVs. In mammalian cells, the produced 25-27 nucleotide long siRNA molecules are processed by endogenous Dicer into 21-nucleotide siRNAs and are thus designated as Dicer-substrate siRNAs (DsiRNAs). We evaluated the efficacy of the above DsiRNA swarm at preventing IAV infections in human primary monocyte-derived macrophages and dendritic cells. The replication of different IAV strains, including avian influenza H5N1 and H7N9 viruses, was significantly inhibited by pre-transfection of the cells with the IAV-specific DsiRNA swarm. Up to 7 orders of magnitude inhibition of viral RNA expression was observed, which led to a dramatic inhibition of IAV protein synthesis and virus production. The IAV-specific DsiRNA swarm inhibited virus replication directly through the RNA interference pathway although a weak induction of innate interferon responses was detected. Our results provide direct evidence for the feasibility of the siRNA strategy and the potency of DsiRNA swarms in the prevention and treatment of influenza, including the highly pathogenic avian influenza viruses.IMPORTANCE In spite of the enormous amount of research, influenza virus is still one of the major challenges for medical virology due to its capacity to generate new variants, which potentially lead to severe epidemics and pandemics. We demonstrated here that a swarm of small interfering RNA (siRNA) molecules, including more than one hundred different antiviral RNA molecules targeting the most conserved regions of influenza A virus genome, could efficiently inhibit the replication of all tested avian and seasonal influenza A variants in human primary monocyte-derived macrophages and dendritic cells. The wide antiviral spectrum makes the virus-specific siRNA swarm a potentially efficient treatment modality against both avian and seasonal influenza viruses.
PMID: 30463970 DOI: 10.1128/JVI.01916-18