The efficacy of lipid-encapsulated, chemically modified short interfering RNA (siRNA) targeted to hepatitis B virus (HBV) was examined in an in vivo mouse model of HBV replication. Stabilized siRNA targeted to the HBV RNA was incorporated into a specialized liposome to form a stable nucleic-acid-lipid particle (SNALP) and administered by intravenous injection into mice carrying replicating HBV. The improved efficacy of siRNA-SNALP compared to unformulated siRNA correlates with a longer half-life in plasma and liver. Three daily intravenous injections of 3 mg/kg/day reduced serum HBV DNA >1.0 log10. The reduction in HBV DNA was specific, dose-dependent and lasted for up to 7 d after dosing. Furthermore, reductions were seen in serum HBV DNA for up to 6 weeks with weekly dosing. The advances demonstrated here, including persistence of in vivo activity, use of lower doses and reduced dosing frequency are important steps in making siRNA a clinically viable therapeutic approach.
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Chiu, Y.L. & Rana, T.M. siRNA function in RNAi: a chemical modification analysis. RNA 9, 1034–1048 (2003).
Layzer, J.M. et al. In vivo activity of nuclease-resistant siRNAs. RNA 10, 766–771 (2004).
Morrissey, D.V. et al. Activity of stabilized siRNAs in a mouse model of HBV replication. Hepatology 41, 1349–1356 (2005).
Ambegia, E. et al. Stabilized plasmid–lipid particles containing PEG-diacylglycerols exhibit extended circulation lifetimes and tumor selective gene expression. Biochim. Biophys. Acta 1669, 155–163 (2005).
Bally, M., Mayer, L., Hope, M. & Nayar, R. Pharmacodynamics of liposomal drug carriers: methodological considerations. in Liposome Technology, vol. III, 27–41, (CRC Press, London, 1993).
Fenske, D.B., MacLachlan, I. & Cullis, P. Long circulating vectors for the systemic delivery of genes. Curr. Opin. Mol. Ther. 3, 153–158 (2001).
Fenske, D.B., MacLachlan, I. & Cullis, P.R. Stabilized plasmid-lipid particles: a systemic gene therapy vector. Methods Enzymol. 346, 36–71 (2002).
Judge, A. et al. Sequence-dependent stimulation of the mammalian innate immune response by synthetic siRNA. Nat. Biotechnol. 23, 457–462 (2005).
Hornung, V. et al. Sequence-specific potent induction of IFN-a by short interfering RNA in plasmacytoid dendritic cells through TLR7. Nat. Med. 11, 263–270 (2005).
Liu, F., Song, Y. & Liu, D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 6, 1258–1266 (1999).
Yang, P.L., Althage, A., Chung, J. & Chisari, F.V. Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc. Natl. Acad. Sci. USA 99, 13825–13830 (2002).
Samuel, C.E. Antiviral actions of interferons. Clin. Microbiol. Rev. 14, 778–809 (2001).
Kim, D.H. et al. Interferon induction by siRNAs and ssRNAs synthesized by phage polymerase. Nat. Biotechnol. 22, 321–325 (2004).
Soutschek, J. et al. Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. Nature 432, 173–178 (2004).
Wincott, F. et al. Synthesis, deprotection, analysis and purification of RNA and ribozymes. Nucleic Acids Res. 23, 2677–2684 (1995).
Wheeler, J.J. et al. Stabilized plasmid-lipid particles: construction and characterization. Gene Ther. 6, 271–281 (1999).
Jeffs, L. et al. A scalable, extrusion free method for efficient liposomal encapsulation of plasmid DNA. Pharm. Res. 22, 362–372 (2005).
Sandberg, J.A. et al. Pharmacokinetics and tolerability of an antiangiogenic ribozyme (ANGIOZYME) in healthy volunteers. J. Clin. Pharmacol. 40, 1462–1469 (2000).
Buckwold, V.E., Xu, Z., Chen, M., Yen, T.S. & Ou, J.H. Effects of a naturally occurring mutation in the hepatitis B virus basal core promoter on precore gene expression and viral replication. J. Virol. 70, 5845–5851 (1996).
The authors would like to thank Roger Aitchison for statistical analysis, and Adrianna Wells for manuscript preparation.
The authors declare no competing financial interests.
SNALP Lipids. (PDF 70 kb)
SNALP Structure. (PDF 685 kb)
In vivo localization of SNALP formulated Cy3-labeled siRNA in mouse livers. (PDF 481 kb)
Chemically stabilized HBV siRNA do not induce local interferon production in the liver. (PDF 81 kb)
Chemically stabilized HBV siRNA do not induce local interferon-beta production in the liver. (PDF 90 kb)
NOD.scid mice generate robust Interferon to unmodified siRNA. (PDF 47 kb)
About this article
Cite this article
Morrissey, D., Lockridge, J., Shaw, L. et al. Potent and persistent in vivo anti-HBV activity of chemically modified siRNAs. Nat Biotechnol 23, 1002–1007 (2005). https://doi.org/10.1038/nbt1122
Nature Reviews Drug Discovery (2021)
Nature Reviews Drug Discovery (2020)
Signal Transduction and Targeted Therapy (2020)
Science China Life Sciences (2020)
Journal of Cardiovascular Translational Research (2020)