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

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...

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Veröffentlicht in:Antimicrobial agents and chemotherapy 2014-05, Vol.58 (5), p.2816-2824
Hauptverfasser: KHANTASUP, Kannika, KOPERMSUB, Phikulthong, CHAICHOUN, Kridsada, DHARAKUL, Tararaj
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container_issue 5
container_start_page 2816
container_title Antimicrobial agents and chemotherapy
container_volume 58
creator KHANTASUP, Kannika
KOPERMSUB, Phikulthong
CHAICHOUN, Kridsada
DHARAKUL, Tararaj
description 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.
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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.</description><identifier>ISSN: 0066-4804</identifier><identifier>EISSN: 1098-6596</identifier><identifier>DOI: 10.1128/AAC.02768-13</identifier><identifier>PMID: 24614365</identifier><identifier>CODEN: AACHAX</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Animals ; Antibiotics. Antiinfectious agents. 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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.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>24614365</pmid><doi>10.1128/AAC.02768-13</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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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
title Targeted Small Interfering RNA-Immunoliposomes as a Promising Therapeutic Agent against Highly Pathogenic Avian Influenza A (H5N1) Virus Infection
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