Targeted Small Interfering RNA-Immunoliposomes as a Promising Therapeutic Agent against Highly Pathogenic Avian Influenza A (H5N1) Virus Infection

• Published in: Antimicrobial agents and chemotherapy Vol. 58; no. 5; pp. 2816 - 2824
• Main Authors: KHANTASUP, Kannika, KOPERMSUB, Phikulthong, CHAICHOUN, Kridsada, DHARAKUL, Tararaj
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.05.2014
• Subjects: Animals; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral Agents; Antiviral Agents - chemistry; Antiviral Agents - pharmacology; Avian influenza virus; Biological and medical sciences; Cell Line; Cell Survival; Dogs; Flow Cytometry; Human viral diseases; Infectious diseases; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H5N1 Subtype - drug effects; Influenza A Virus, H5N1 Subtype - pathogenicity; Influenza virus; Liposomes; Liposomes - chemistry; Medical sciences; Pharmacology. Drug treatments; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; RNA, Small Interfering - chemistry; RNA, Small Interfering - genetics; Single-Chain Antibodies; Single-Chain Antibodies - chemistry; Viral diseases; Viral diseases of the respiratory system and ent viral diseases; RNA interference; Targeting; Orthomyxoviridae; siRNA; Infection; Virus; Avian influenza; Influenzavirus A(H5N1); Gene silencing; Target; Influenzavirus A; Treatment; Influenza A virus; Viral disease; Pathogenic; Immunoliposome
• ISSN: 0066-4804; 1098-6596
• DOI: 10.1128/AAC.02768-13

This study describes a proof-of-concept study on the use of small interfering RNA (siRNA)-immunoliposomes as a therapeutic agent against H5N1 influenza virus infection. siRNA specific for influenza virus nucleoprotein (NP) mRNA was employed as the key antiviral agent to inhibit viral replication in this study. A humanized single-chain Fv antibody (huscFv) against the hemagglutinin (HA) of H5N1 highly pathogenic avian influenza virus (HPAI) was used as the targeting molecule to HA of H5N1 virus, which is abundantly expressed on the surface of infected cells (the HA target cells). The huscFv was applied to cationic polyethylene glycol-conjugated 3β-[N-(N',N'-dimethylaminoethane) carbamoyl] cholesterol-dioleoylphosphatidyl ethanolamine (PEGylated DC-Chol-DOPE) liposomes to generate immunoliposomes for siRNA delivery. The immunoliposomes were shown to specifically bind HA-expressing Sf9 cells and demonstrated enhanced siRNA transfection efficiency. The siRNA transfection efficiency was significantly reduced after preincubation of the HA target cells with an excess amount of free huscFv. These results therefore demonstrated that the enhanced siRNA delivery by use of immunoliposomes was mediated via targeting by huscFv. Furthermore, the siRNA silencing effect was more pronounced when the immunoliposomes were administered 6 to 12 h post-H5N1 infection in MDCK cells compared with the nontargeted liposomes. This proof-of-concept study may contribute to the future design and development of an siRNA delivery system for combating viral infectious diseases in humans.