Efficient inhibition of hepatitis B virus replication in vivo, using polyethylene glycol-modified adenovirus vectors.
Achieving safe delivery of anti-hepatitis B virus (HBV) RNA interference (RNAi) effectors is an important objective of this gene-silencing technology. Adenoviruses (Ads) have a natural tropism for the liver after systemic administration, and are useful for delivery of expressed anti-HBV RNAi sequences. However, a drawback of Ad vectors is diminished efficacy and toxicity that results from stimulation of innate and adaptive immunity. To attenuate these effects we used monomethoxy polyethylene glycol-succinimidyl propionate (mPEG-SPA) to modify first-generation vectors that express an anti-HBV RNAi effector. Efficient hepatocyte transduction and knockdown of HBV replication were achieved after intravenous administration of 5 x 10(9) PEGylated or native recombinant Ads to HBV transgenic mice. After the first injection, circulating HBV viral particle equivalents (VPEs) remained low for 3 weeks and began to increase after 5 weeks. A second dose of PEGylated anti-HBV Ad caused a less sustained decrease in circulating VPEs, but no silencing after a second dose was observed in animals treated with unmodified vector. Release of inflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), interferon-gamma, interleukin-6, and tumor necrosis factor-alpha, was elevated in animals receiving unmodified vectors. However, only a modest increase in MCP-1 was observed in mice that received a second dose of PEG Ads. Also, polymer-conjugated vectors induced a weaker adaptive immune response and were less hepatotoxic than their unmodified counterparts. Collectively, these observations show that PEG modification of Ads expressing RNAi effectors improves their potential for therapeutic application against HBV infection.
Department of Molecular Medicine and Hematology, University of the Witwatersrand, Johannesburg, South Africa.
Carol
C
Crowther
Abdullah
A
Ely
Judith
J
Hornby
Steven
S
Mufamadi
Felix
F
Salazar
Patricia
P
Marion
Patrick
P
Arbuthnot
D000256:Adenoviridae / Q000235:genetics*; D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D015152:Blotting, Northern; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006515:Hepatitis B virus / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D011092:Polyethylene Glycols / Q000737:chemistry*; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology*
D016207:Cytokines; D034741:RNA, Small Interfering; D011092:Polyethylene Glycols; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase