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Suppression of Apoptosis in the Liver by Systemic and Local Delivery of Small-Interfering RNAs

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Target Discovery and Validation Reviews and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 361))

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Abstract

RNA interference (RNAi) is a sequence-specific gene-silencing mechanism triggered by double-stranded RNA. RNAi was shown to allow transient or stable knockdown of gene expression in a broad range of species and has been used successfully for functional genomic screens in mammalian cells and Caenorhabditis elegans. Standard therapeutic use of RNAi in clinical settings in humans has been hampered by the lack of effective methods to deliver the small-interfering RNAs (siRNAs) or short-hairpin RNA expression vectors into the diseased organs. In mice, systemic delivery of siRNAs by hydrodynamic intravascular injection leads to highly efficient uptake of siRNAs into the liver. Several groups demonstrated therapeutic use of RNAi in mouse models of acute liver failure or hepatitis B virus replication. This chapter will focus on the technical background of hydrodynamic and portal vein delivery techniques in mice and will give practical guidance for using these techniques for siRNA delivery into the liver.

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References

  1. Paddison, P. J., Silva, J. M., Conklin, D. S., et al. (2004) A resource for large-scale RNA-interference-based screens in mammals. Nature 428, 427–431.

    Article  CAS  PubMed  Google Scholar 

  2. Berns, K., Hijmans, E. M., Mullenders, J., et al. (2004) A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428, 431–437.

    Article  CAS  PubMed  Google Scholar 

  3. Brummelkamp, T. R., Nijman, S. M., Dirac, A. M., and Bernards, R. (2003) Loss of the cylindromatosis tumour suppressor inhibits apoptosis by activating NF-kappaB. Nature 424, 797–801.

    Article  CAS  PubMed  Google Scholar 

  4. Zender, L., Hutker, S., Liedtke, C., et al. (2003) Caspase 8 small interfering RNA prevents acute liver failure in mice. Proc. Natl. Acad. Sci. USA 100, 7797–7802.

    Article  CAS  PubMed  Google Scholar 

  5. Song, E., Lee, S. K., Wang, J., et al. (2003) RNA interference targeting Fas protects mice from fulminant hepatitis. Nat. Med. 9, 347–351.

    Article  CAS  PubMed  Google Scholar 

  6. Klein, C., Bock, C. T., Wedemeyer, H., et al. (2003) Inhibition of hepatitis B virus replication in vivo by nucleoside analogues and siRNA. Gastroenterology 125, 9–18.

    Article  CAS  PubMed  Google Scholar 

  7. McCaffrey, A. P., Nakai, H., Pandey, K., et al. (2003) Inhibition of hepatitis B virus in mice by RNA interference. Nat. Biotechnol. 21, 639–644.

    Article  CAS  PubMed  Google Scholar 

  8. Bertrand, J. R., Pottier, M., Vekris, A., Opolon, P., Maksimenko, A., and Malvy, C. (2002) Comparison of antisense oligonucleotides and siRNAs in cell culture and in vivo. Biochem. Biophys. Res. Commun. 296, 1000–1004.

    Article  CAS  PubMed  Google Scholar 

  9. Danko, I., Williams, P., Herweijer, H., et al. (1997) High expression of naked plasmid DNA in muscles of young rodents. Hum. Mol. Genet. 6, 1435–1443.

    Article  CAS  PubMed  Google Scholar 

  10. Hickman, M. A., Malone, R. W., Lehmann-Bruinsma, K., et al. (1994) Gene expression following direct injection of DNA into liver. Hum. Gene Ther. 5, 1477–1483.

    Article  CAS  PubMed  Google Scholar 

  11. Hartikka, J., Sukhu, L., Buchner, C., et al. (2001) Electroporation-facilitated delivery of plasmid DNA in skeletal muscle: plasmid dependence of muscle damage and effect of poloxamer 188. Mol. Ther. 4, 407–415.

    Article  CAS  PubMed  Google Scholar 

  12. Somiari, S., Glasspool-Malone, J., Drabick, J. J., et al. (2000) Theory and in vivo application of electroporative gene delivery. Mol. Ther. 2, 178–187.

    Article  CAS  PubMed  Google Scholar 

  13. Budker, V., Zhang, G., Knechtle, S., and Wolff, J. A. (1996) Naked DNA delivered intraportally expresses efficiently in hepatocytes. Gene Ther. 3, 593–598.

    CAS  PubMed  Google Scholar 

  14. Zhang, G., Budker, V., and Wolff, J. A. (1999) High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. Hum. Gene Ther. 10, 1735–1737.

    Article  CAS  PubMed  Google Scholar 

  15. McCaffrey, A. P., Meuse, L., Pham, T. T., Conklin, D. S., Hannon, G. J., and Kay, M. A. (2002) RNA interference in adult mice. Nature 418, 38–39.

    Article  CAS  PubMed  Google Scholar 

  16. Song, E., Lee, S. K., Wang, J., et al. (2003) RNA interference targeting Fas protects mice from fulminant hepatitis. Nat. Med. 9, 347–351.

    Article  CAS  PubMed  Google Scholar 

  17. Zender, L., Hutker, S., Liedtke, C., et al. (2003) Caspase 8 small interfering RNA prevents acute liver failure in mice. Proc. Natl. Acad. Sci. USA 100, 7797–7802.

    Article  CAS  PubMed  Google Scholar 

  18. Budker, V., Budker, T., Zhang, G., Subbotin, V., Loomis, A., and Wolff, J. A. (2000) Hypothesis: naked plasmid DNA is taken up by cells in vivo by a receptor-mediated process. J. Gene Med. 2, 76–88.

    Article  CAS  PubMed  Google Scholar 

  19. Kobayashi, N., Kuramoto, T., Yamaoka, K., Hashida, M., and Takakura, Y. (2001) Hepatic uptake and gene expression mechanisms following intravenous administration of plasmid DNA by conventional and hydrodynamics-based procedures, J. Pharmacol. Exp. Ther. 297, 853–860.

    CAS  PubMed  Google Scholar 

  20. O’Grady, J. G., Schalm, S. W., and Williams, R. (1993) Acute liver failure: redefining the syndromes. Lancet 342, 273–275.

    Article  PubMed  Google Scholar 

  21. Plevris, J. N., Schina, M., and Hayes, P. C. (1998) Review article: the management of acute liver failure. Aliment. Pharmacol. Ther. 12, 405–418.

    Article  CAS  PubMed  Google Scholar 

  22. Bernal, W. (2003) Changing patterns of causation and the use of transplantation in the United Kingdom. Semin. Liver Dis. 23, 227–237.

    Article  PubMed  Google Scholar 

  23. Jaeschke, H., Gores, G. J., Cederbaum, A. I., Hinson, J. A., Pessayre, D., and Lemasters, J. J. (2002) Mechanisms of hepatotoxicity. Toxicol. Sci. 65, 166–176.

    Article  CAS  PubMed  Google Scholar 

  24. Adams, J. M. and Cory, S. (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281, 1322–1326.

    Article  CAS  PubMed  Google Scholar 

  25. Ashkenazi, A. and Dixit, V. M. (1998) Death receptors: signaling and modulation, Science 281, 1305–1308.

    Article  CAS  PubMed  Google Scholar 

  26. Evan, G. and Littlewood, T. (1998) A matter of life and cell death. Science 281, 1317–1322.

    Article  CAS  PubMed  Google Scholar 

  27. Green, D. R. and Reed, J. C. (1998) Mitochondria and apoptosis. Science 281, 1309–1312.

    Article  CAS  PubMed  Google Scholar 

  28. Galle, P. R., Hofmann, W. J., Walczak, H., et al. (1995) Involvement of the CD95 (APO-1/Fas) receptor and ligand in liver damage. J. Exp. Med. 182, 1223–1230.

    Article  CAS  PubMed  Google Scholar 

  29. Strand, S., Hofmann, W. J., Grambihler, A., et al. (1998) Hepatic failure and liver cell damage in acute Wilson’s disease involve CD95 (APO-1/Fas) mediated apoptosis. Nat. Med. 4, 588–593.

    Article  CAS  PubMed  Google Scholar 

  30. Kuhnel, F., Zender, L., Paul, Y., et al. (2000) NFkappaB mediates apoptosis through transcriptional activation of Fas (CD95) in adenoviral hepatitis. J. Biol. Chem. 275, 6421–6427.

    Article  CAS  PubMed  Google Scholar 

  31. Faubion, W. A., Guicciardi, M. E., Miyoshi, H., et al. (1999) Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas. J. Clin. Invest. 103, 137–145.

    Article  CAS  PubMed  Google Scholar 

  32. Feldstein, A. E., Canbay, A., Angulo, P., et al. (2003) Hepatocyte apoptosis and fas expression are prominent features of human nonalcoholic steatohepatitis. Gastroenterology 125, 437–443.

    Article  PubMed  Google Scholar 

  33. Yin, X. M. and Ding, W. X. (2003) Death receptor activation-induced hepatocyte apoptosis and liver injury. Curr. Mol. Med. 3, 491–508.

    Article  CAS  PubMed  Google Scholar 

  34. Zender, L., Hutker, S., Liedtke, C., et al. (2003) Caspase 8 small interfering RNA prevents acute liver failure in mice. Proc. Natl. Acad. Sci. USA 100, 7797–7802.

    Article  CAS  PubMed  Google Scholar 

  35. Zhang, H., Cook, J., Nickel, J., et al. (2000) Reduction of liver Fas expression by an antisense oligonucleotide protects mice from fulminant hepatitis. Nat. Biotechnol. 18, 862–867.

    Article  CAS  PubMed  Google Scholar 

  36. Mundt, B., Kuhnel, F., Zender, L., et al. (2003) Involvement of TRAIL and its receptors in viral hepatitis. FASEB J. 17, 94–96.

    CAS  PubMed  Google Scholar 

  37. Streetz, K. L., Luedde, T., Manns, M. P., and Trautwein, C. (2000) Interleukin 6 and liver regeneration. Gut 47, 309–312.

    Article  CAS  PubMed  Google Scholar 

  38. Seglen, P. O., Schwarze, P. E., and Saeter, G. (1986) Changes in cellular ploidy and autophagic responsiveness during rat liver carcinogenesis. Toxicol. Pathol. 14, 342–348.

    Article  CAS  PubMed  Google Scholar 

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© 2007 Humana Press Inc.

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Zender, L., Kubicka, S. (2007). Suppression of Apoptosis in the Liver by Systemic and Local Delivery of Small-Interfering RNAs. In: Sioud, M. (eds) Target Discovery and Validation Reviews and Protocols. Methods in Molecular Biology™, vol 361. Humana Press. https://doi.org/10.1385/1-59745-208-4:217

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  • DOI: https://doi.org/10.1385/1-59745-208-4:217

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-890-4

  • Online ISBN: 978-1-59745-208-3

  • eBook Packages: Springer Protocols

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