Abstract
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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
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).
Acknowledgements
The authors would like to thank Roger Aitchison for statistical analysis, and Adrianna Wells for manuscript preparation.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
SNALP Lipids. (PDF 70 kb)
Supplementary Fig. 2
SNALP Structure. (PDF 685 kb)
Supplementary Fig. 3
In vivo localization of SNALP formulated Cy3-labeled siRNA in mouse livers. (PDF 481 kb)
Supplementary Fig. 4
Chemically stabilized HBV siRNA do not induce local interferon production in the liver. (PDF 81 kb)
Supplementary Fig. 5
Chemically stabilized HBV siRNA do not induce local interferon-beta production in the liver. (PDF 90 kb)
Supplementary Fig. 6
NOD.scid mice generate robust Interferon to unmodified siRNA. (PDF 47 kb)
Rights and permissions
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nbt1122
This article is cited by
-
Oligonucleotide therapeutics and their chemical modification strategies for clinical applications
Journal of Pharmaceutical Investigation (2024)
-
Biomembrane-derived nanoplexes for SiRNAs-pioneer innovation in delivery to lung adenocarcinoma
Journal of Nanoparticle Research (2024)
-
Engineering siRNA therapeutics: challenges and strategies
Journal of Nanobiotechnology (2023)
-
A practical approach to RNA interference for studying gene function in a refractory social insect (on a limited budget)
Insectes Sociaux (2023)
-
Innate immune regulations and various siRNA modalities
Drug Delivery and Translational Research (2023)