Comparison of Influenza a Virus Inhibition in Vitro by Sirna Complexes With Chitosan Derivatives, Polyethyleneimine and Hybrid Polyarginine-Inorganic Microcapsules

Abstract

Anti-influenza drugs and vaccines have a limited effect due to the high mutation rate of virus genome. The direct impact on the conservative virus genome regions should significantly improve therapeutic effectiveness. The RNA interference mechanism (RNAi) is one of the modern approaches used to solve this problem. In this work, we have investigated the antiviral activity of small interfering RNA (siRNA) against the influenza A/PR/8/34 (H1N1), targeting conserved regions of NP and PA. Polycations were used for intracellular siRNA delivery: chitosan’s derivatives (methylglycol and quaternized chitosan), polyethyleneimine, lipofectamine, and hybrid organic/non-organic microcapsules. A comparative study of these delivery systems with fluorescent labeled siRNA was conducted. The antiviral activity of three small interfering RNAs targeting the NP (NP-717, NP-1496) and PA (PA-1630) influenza A viruses genes was demonstrated, depending on the chosen carrier. The most effective intracellular delivery and antiviral activity were observed for hybrid microcapsules.

Progress in non-viral localized delivery of siRNA therapeutics for pulmonary diseases

“…Hybrid organic-inorganic microcarriers 2. siRNA complexed with pH-responsive peptides 3. Chitosan/siRNA nanoparticles In vitro; in vitro; intranasal Effective silencing reported in both studies; pH responsive peptides improved endolysosomes escape of siRNA; nasal delivery of siRNA particles significantly protected mice from a lethal influenza virus challenge 70 , 71 , 72 NP and PA Methylglycolchitosan, quaternized chitosan, polyethyleneimine, and hybrid microcapsules based on polyarginine with inorganic components In vitro Hybrid capsules were identified as the best carrier with the most effective intracellular delivery and highest antiviral activity 73 NP , NS and PA cocktail siRNA siRNA encapsulation in a hybrid microcapsule made of polyarginine-dextran sulfate polyelectrolytes coated by SiO 2 In vitro Intracellular uptake was more efficient than commercial liposome/PEI-based transfection reagent and reported effective gene inhibition in several influenza A virus subtypes 74 NS1 Lipofectamine 2000 transfection agent Intranasal Reduced virus-induced death rate in infected chicken by up to 87.5% with significantly lower viral titers and plaque formation in the chicken lungs 75 Tuberculosis BFL-1/A1 Naked siRNA In vitro siRNA inhibition induced apoptosis in both Mtb-infected THP-1 cells and macrophages and enhanced phagosome-lysosome functions 76 RAB22a Naked siRNA In vitro Rab22a knockdown disinhibited Rab7 acquisition and phagosome maturation in Mtb-infected macrophages 77 TGFß Naked siRNA Inhalation Mice treated with TGF-β siRNA showed increased antimicrobial mediators ( e.g. , NO and iNOS) and significantly reduced bacterial load 78 XCL1 Naked siRNA Intratracheal Observed a transient but strong modulation of the production of XCL1 in the lungs and significantly reduced IFN-r secreting T cells 79 …”

“… 72 formulated chitosan/siRNA nanoparticles to inhibit influenza virus replication in BALB/c mice after intranasal delivery. Other materials such as hybrid polyarginine-inorganic microcapsules have also been developed for siRNA delivery against influenza 73 .…”

Layer-by-Layer technique as a versatile tool for gene delivery applications

“…Figure 3(a) (bottom middle) reproduced form [78] with permission of Royal Society of Chemistry. Figure 3(a) (top right) reproduced form [79] with permission of Wiley publishing group. Figure 3(a) (bottom right) reproduced form [80] with permission of American Chemistry Society Publications.…”