tetano
Editor, Senior Moderator
Antiviral Res. 2018 Aug 6. pii: S0166-3542(17)30732-5. doi: 10.1016/j.antiviral.2018.08.003. [Epub ahead of print]
[h=1]Inhibition of influenza A virus by mixed siRNAs, targeting the PA, NP, and NS genes, delivered by hybrid microcarriers.[/h] Brodskaia AV[SUP]1[/SUP], Timin AS[SUP]2[/SUP], Gorshkov AN[SUP]3[/SUP], Muslimov AR[SUP]4[/SUP], Bondarenko AB[SUP]5[/SUP], Tarakanchikova YV[SUP]6[/SUP], Zabrodskaya YA[SUP]7[/SUP], Baranovskaya IL[SUP]8[/SUP], Il'inskaja EV[SUP]9[/SUP], Sakhenberg EI[SUP]3[/SUP], Sukhorukov GB[SUP]10[/SUP], Vasin AV[SUP]11[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] In the present study, a highly effective carrier system has been developed for the delivery of antiviral siRNA mixtures. The developed hybrid microcarriers, made of biodegradable polymers and SiO[SUB]2[/SUB] nanostructures, more efficiently mediate cellular uptake of siRNA than commercially available liposome-based reagents and polyethyleneimine (PEI); they also demonstrate low in vitro toxicity and protection of siRNA from RNase degradation. A series of siRNA designs (targeting the most conserved regions of three influenza A virus (IAV) genes: NP, NS, and PA) were screened in vitro using RT-qPCR, ELISA analysis, and hemagglutination assay. Based on the results of screening, the three most effective siRNAs (PA-1630, NP-717, and NS-777) were selected for in situ encapsulation into hybrid microcarriers. It was revealed that pre-treatment of cells with a mixture of PA-1630, NP-717, and NS-777 siRNAs, delivered by hybrid microcarriers, provided stronger inhibition of viral M1 mRNA expression and control of NP protein level, after viral infection, than single pre-treatment by any of three encapsulated siRNAs used in the study. Moreover, the effective inhibition of replication in several IAV subtypes (H1N1, H1N1pdm, H5N2, and H7N9) using a cocktail of the three selected siRNAs, delivered by our hybrid capsules to the cells, was achieved. In conclusion, we have developed a proof-of-principle which shows that our hybrid microcarrier technology (utilizing a therapeutic siRNA cocktail) may represent a promising approach in anti-influenza therapy.
[h=4]KEYWORDS:[/h] Hybrid microcontainers; Influenza A virus; RNAi therapy; siRNA delivery
PMID: 30092251 DOI: 10.1016/j.antiviral.2018.08.003
[h=1]Inhibition of influenza A virus by mixed siRNAs, targeting the PA, NP, and NS genes, delivered by hybrid microcarriers.[/h] Brodskaia AV[SUP]1[/SUP], Timin AS[SUP]2[/SUP], Gorshkov AN[SUP]3[/SUP], Muslimov AR[SUP]4[/SUP], Bondarenko AB[SUP]5[/SUP], Tarakanchikova YV[SUP]6[/SUP], Zabrodskaya YA[SUP]7[/SUP], Baranovskaya IL[SUP]8[/SUP], Il'inskaja EV[SUP]9[/SUP], Sakhenberg EI[SUP]3[/SUP], Sukhorukov GB[SUP]10[/SUP], Vasin AV[SUP]11[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] In the present study, a highly effective carrier system has been developed for the delivery of antiviral siRNA mixtures. The developed hybrid microcarriers, made of biodegradable polymers and SiO[SUB]2[/SUB] nanostructures, more efficiently mediate cellular uptake of siRNA than commercially available liposome-based reagents and polyethyleneimine (PEI); they also demonstrate low in vitro toxicity and protection of siRNA from RNase degradation. A series of siRNA designs (targeting the most conserved regions of three influenza A virus (IAV) genes: NP, NS, and PA) were screened in vitro using RT-qPCR, ELISA analysis, and hemagglutination assay. Based on the results of screening, the three most effective siRNAs (PA-1630, NP-717, and NS-777) were selected for in situ encapsulation into hybrid microcarriers. It was revealed that pre-treatment of cells with a mixture of PA-1630, NP-717, and NS-777 siRNAs, delivered by hybrid microcarriers, provided stronger inhibition of viral M1 mRNA expression and control of NP protein level, after viral infection, than single pre-treatment by any of three encapsulated siRNAs used in the study. Moreover, the effective inhibition of replication in several IAV subtypes (H1N1, H1N1pdm, H5N2, and H7N9) using a cocktail of the three selected siRNAs, delivered by our hybrid capsules to the cells, was achieved. In conclusion, we have developed a proof-of-principle which shows that our hybrid microcarrier technology (utilizing a therapeutic siRNA cocktail) may represent a promising approach in anti-influenza therapy.
[h=4]KEYWORDS:[/h] Hybrid microcontainers; Influenza A virus; RNAi therapy; siRNA delivery
PMID: 30092251 DOI: 10.1016/j.antiviral.2018.08.003