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Delivery of Adenoviral DNA to Mouse Liver

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Gene Delivery to Mammalian Cells

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 246))

Abstract

The liver represents a major target organ for gene delivery owing to its high biosynthetic capacity and access to the bloodstream. Adenoviral vectors are highly efficient gene-transfer vehicles, making them among the most promising systems for in vivo gene transfer to the liver. Following intravenous administration of adenoviral vectors to a variety of mammalian models, including mice, dogs, and monkeys, hepatocytes are efficiently transduced (1). Several delivery methods to the liver have been described, including portal vein (2–4), hepatic artery (3,5), and peripheral vein infusions (6). This chapter describes the simple, nonsurgical method of intravenous (iv) administration of adenoviral vectors in mice, and an immunohistochemical method to qualitatively evaluate liver transduction efficiency following delivery of an adenoviral vector encoding a β-galactosidase (β-gal) marker gene. Additionally, several alternative methods to verify efficient liver transduction are introduced.

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References

  1. Connelly, S. (1999) Adenoviral vectors for liver-directed gene therapy. Curt Opin. Mol. Ther. 1, 565–572.

    CAS  Google Scholar 

  2. Vrancken Peeters, M. J., Perkins, A. L., and Kay, M. A. (1996) Method for multiple portal vein infusions in mice: quantitation of adenovirus-mediated hepatic gene transfer. Biotechniques 20, 278–285.

    PubMed  CAS  Google Scholar 

  3. Gerolami, R., Cardoso, J., Bralet, M. P., Cuenod, C. A., Clement, O., Tran, P. L., and Brechot, C. (1998) Enhanced in vivo adenovirus-mediated gene transfer to rat hepatocarcinomas by selective administration into the hepatic artery. Gene Ther. 5, 896–904.

    Article  PubMed  CAS  Google Scholar 

  4. Cichon, G., Schmidt, H. H., Benhidjeb, T., Loser, P., Ziemer, S., Haas, R., et al. (1999) Intravenous administration of recombinant adenoviruses causes thrombocytopenia, anemia and erythroblastosis in rabbits. J. Gene Med. 1, 360–371.

    Article  PubMed  CAS  Google Scholar 

  5. Maron, D. J., Tada, H., Moscioni, A. D., Tazelaar, J., Fraker, D. L., Wilson, J. M., and Spitz, F.R. (2001) Intra-arterial delivery of a recombinant adenovirus does not increase gene transfer to tumor cells in a rat model of metastatic colorectal carcinoma. Mol. Ther. 4, 29–35.

    Article  PubMed  CAS  Google Scholar 

  6. Smith, T. A., Mehaffey, M. G., Kayda, D. B., Saunders, J. M., Yei, S., Trapnell, B. C., et al. (1993) Adenovirus mediated expression of therapeutic plasma levels of human factor IX in mice. Nat. Genet. 5, 397–402.

    Article  PubMed  CAS  Google Scholar 

  7. Yang, Y., Nunes, F. A., Berencsi, K., Furth, E. E., Gonczol, E., and Wilson, J. M. (1994) Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy. Proc. Natl. Acad. Sci. USA 91, 4407–4411.

    Article  PubMed  CAS  Google Scholar 

  8. Yang, Y, Jooss, K. U., Su, Q., Ertl, H. C, and Wilson, J. M. (1996) Immune responses to viral antigens versus transgene product in the elimination of recombinant adenovirus-infected hepatocytes in vivo. Gene Ther. 3, 137–144.

    PubMed  Google Scholar 

  9. Yang, Y. and Wilson, J. M. (1995) Clearance of adenovirus-infected hepatocytes by MHC class I-restricted CD4+ CTLs in vivo. J. Immunol. 155, 2564–2570.

    PubMed  CAS  Google Scholar 

  10. Berkner, K. L. (1988) Development of adenovirus vectors for the expression of heterologous genes. Biotechniques 6, 616–629.

    Article  PubMed  CAS  Google Scholar 

  11. Gorziglia, M. I., Kadan, M. J., Yei, S., Lim, J., Lee, G. M., Luthra, R., and Trapnell, B. C. (1996) Elimination of both E1 and E2 from adenovirus vectors further improves prospects for in vivo human gene therapy. J. Virol. 70, 4173–4178.

    PubMed  CAS  Google Scholar 

  12. Gorziglia, M. I., Lapcevich, C., Roy, S., Kang, Q., Kadan, M., Wu, V., et al. (1999) Generation of an adenovirus vector lacking E1, E2a, E3, and all of E4 except open reading frame 3. J. Virol. 73, 6048–6055.

    PubMed  CAS  Google Scholar 

  13. Morral, N., O’Neal, W., Zhou, H., Langston, C., and Beaudet, A. (1997) Immune responses to reporter proteins and high viral dose limit duration of expression with adenoviral vectors: comparison of E2a wild type and E2a deleted vectors. Hum. Gene Ther. 8, 1275–1286.

    Article  PubMed  CAS  Google Scholar 

  14. Amalfitano, A. and Chamberlain, J. S. (1997) Isolation and characterization of packaging cell lines that coexpress the adenovirus E1, DNA polymerase, and preterminal proteins: implications for gene therapy. Gene Ther. 4, 258–263.

    Article  PubMed  CAS  Google Scholar 

  15. Armentano, D., Zabner, J., Sacks, C., Sookdeo, C. C., Smith, M. P., St George, J. A., et al. (1997) Effect of the E4 region on the persistence of transgene expression from adenovirus vectors. J. Virol. 71, 2408–2416.

    PubMed  CAS  Google Scholar 

  16. Andrews, J. L., Kadan, M. J., Gorziglia, M. I., Kaleko, M., and Connelly, S. (2001) Generation and characterization of E1/E2a/E3/E4-deficient adenoviral vectors encoding human factor VIII. Mol. Ther. 3, 329–336.

    Article  PubMed  CAS  Google Scholar 

  17. Brough, D. E., Hsu, C., Kulesa, V. A., Lee, G. M., Cantolupo, L. J., Lizonova, A., and Kovesdi, I. (1997) Activation of transgene expression by early region 4 is responsible for a high level of persistent transgene expression from adenovirus vectors in vivo. J. Virol. 71, 9206–9213.

    PubMed  CAS  Google Scholar 

  18. Lusky, M., Grave, L., Dieterle, A., Dreyer, D., Christ, M., Ziller, C., et al. (1999) Regulation of adenovirus-mediated transgene expression by the viral E4 gene products: requirement for E4 ORF3. J. Virol. 73, 8308–8319.

    PubMed  CAS  Google Scholar 

  19. Christ, M., Louis, B., Stoeckel, F., Dieterle, A., Grave, L., Dreyer, D., et al. (2000) Modulation of the inflammatory properties and hepatotoxicity of recombinant adenovirus vectors by the viral E4 gene products. Hum. Gene Ther. 11, 415–427.

    Article  PubMed  CAS  Google Scholar 

  20. Morral, N., O’Neal, W., Rice, K., Leland, M., Kaplan, J., Piedra, P. A., et al. (1999) Administration of helper-dependent adenoviral vectors and sequential delivery of different vector serotype for long-term liver-directed gene transfer in baboons. Proc. Natl. Acad. Sci. USA 96, 12816–12821.

    Article  PubMed  CAS  Google Scholar 

  21. Sandig, V., Youil, R., Bett, A. J., Franlin, L. L., Oshima, M., Maione, D., et al. (2000) Optimization of the helper-dependent adenovirus system for production and potency in vivo. Proc. Natl. Acad. Sci. USA 97, 1002–1007.

    Article  PubMed  CAS  Google Scholar 

  22. Maione, D., Wiznerowicz, M., Delmastro, P., Cortese, R., Ciliberto, G., La Monica, N., and Savino, R. (2000) Prolonged expression and effective readministration of erythropoietin delivered with a fully deleted adenoviral vector. Hum. Gene Ther. 11, 859–868.

    Article  PubMed  CAS  Google Scholar 

  23. Balague, C., Zhou, J., Dai, Y., Alemany, R., Josephs, S. F., Andreason, G., et al. (2000) Sustained high-level expression of full-length human factor VIII and restoration of clotting activity in hemophilic mice using a minimal adenovirus vector. Blood 95, 820–828.

    PubMed  CAS  Google Scholar 

  24. Kochanek, S., Schiedner, G., and Volpers, C. (2001) High-capacity ‘gutless’ adenoviral vectors. Curr. Opin. Mol. Ther. 3, 454–463.

    PubMed  CAS  Google Scholar 

  25. Reddy, P. S., Sakhuja, K., Ganesh, S., Yang, L., Kayda, D., Brann, T., et al. (2002) Sustained human factor VIII expression in hemophilia a mice following systemic delivery of a gutless adenoviral vector. Mol. Ther. 5, 63–73.

    Article  PubMed  CAS  Google Scholar 

  26. Gao, G. P., Yang, Y., and Wilson, J. M. (1996) Biology of adenovirus vectors with E1 and E4 deletions for liver-directed gene therapy. J. Virol. 70, 8934–8943.

    PubMed  CAS  Google Scholar 

  27. Tripathy, S. K., Svensson, E. C., Black, H. B., Goldwasser, E., Margalith, M., Hobart, P. M., and Leiden, J. M. (1996) Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of a plasmid DNA vector. Proc. Natl. Acad. Sci. USA 93, 10876–10880.

    Article  PubMed  CAS  Google Scholar 

  28. Loser, P., Jennings, G. S., Strauss, M., and Sandig, V. (1998) Reactivation of the previously silenced cytomegalovirus major immediate-early promoter in the mouse liver: involvement of NFkappaB. J. Virol. 72, 180–190.

    PubMed  CAS  Google Scholar 

  29. De Geest, B., Van Linthout, S., Lox, M., Collen, D., and Holvoet, P. (2000) Sustained expression of human apolipoprotein A-I after adenoviral gene transfer in C57BL/6 mice: role of apolipoprotein A-I promoter, apolipoprotein A-I introns, and human apolipoprotein E enhancer. Hum. Gene Ther. 11, 101–112.

    Article  PubMed  Google Scholar 

  30. Connelly, S., Gardner, J. M., McClelland, A., and Kaleko, M. (1996) High-level tissue-specific expression of functional human factor VIII in mice. Hum. Gene Ther. 7, 183–195.

    Article  PubMed  CAS  Google Scholar 

  31. Pastore, L., Morral, N., Zhou, H., Garcia, R., Parks, R. J., Kochanek, S., et al. (1999) Use of a liver-specific promoter reduces immune response to the transgene in adenoviral vectors. Hum. Gene Ther. 10, 1773–1781.

    Article  PubMed  CAS  Google Scholar 

  32. Bristol, J. A., Gallo-Penn, A., Andrews, J., Idamakanti, N., Kaleko, M., and Connelly, S. (2001) Adenovirus-mediated factor VIII gene expression results in attenuated anti-factor VIII-specific immunity in hemophilia A mice compared with factor VIII protein infusion. Hum. Gene Ther. 12, 1651–1661.

    Article  PubMed  CAS  Google Scholar 

  33. Jaffe, H. A., Danel, C., Longenecker, G., Metzger, M., Setoguchi, Y., Rosenfeld, M. A., et al. (1992) Adenovirus-mediated in vivo gene transfer and expression in normal rat liver. Nat. Genet. 1, 372–378.

    Article  PubMed  CAS  Google Scholar 

  34. Hackett, N. R., El Sawy, T., Lee, L. Y., Silva, I., O’Leary, J., Rosengart, T. K., and Crystal, R. G. (2000) Use of quantitative TaqMan real-time PCR to track the time-dependent distribution of gene transfer vectors in vivo. Mol. Ther. 2, 649–656.

    Article  PubMed  CAS  Google Scholar 

  35. Kay, M. A., Landen, C. N., Rothenberg, S. R., Taylor, L. A., Leland, F., Wiehle, S., et al. (1994) In vivo hepatic gene therapy: complete albeit transient correction of factor IX deficiency in hemophilia B dogs. Proc. Natl. Acad. Sci. USA 91, 2353–2357.

    Article  PubMed  CAS  Google Scholar 

  36. Connelly, S., Mount, J., Mauser, A., Gardner, J. M., Kaleko, M., McClelland, A., and Lothrop, C. D., Jr. (1996) Complete short-term correction of canine hemophilia A by in vivo gene therapy. Blood 88, 3846–3853.

    PubMed  CAS  Google Scholar 

  37. Fang, B., Wang, H., Gordon, G., Bellinger, D. A., Read, M. S., Brinkhous, K. M., et al. (1996) Lack of persistence of E1-recombinant adenoviral vectors containing a temperature-sensitive E2A mutation in immunocompetent mice and hemophilia B dogs. Gene Ther. 3, 217–222.

    PubMed  CAS  Google Scholar 

  38. Brann, T., Kayda, D., Lyons, R. M., Shirley, P., Roy, S., Kaleko, M., and Smith, T. (1999) Adenoviral vector-mediated expression of physiologic levels of human factor VIII in nonhuman primates. Hum. Gene Ther. 10, 2999–3011.

    Article  PubMed  CAS  Google Scholar 

  39. Andrews, J. L., Shirley, P. S., Iverson, W. O., Sherer, A. D., Markovits, J. E., King, L., et al. (2002) Evaluation of the duration of human factor VIII expression in non-human primates following systemic delivery of an adenoviral vector. Hum. Gene Ther. 13, 1331–1336.

    Article  PubMed  CAS  Google Scholar 

  40. Michou, A. I., Santoro, L., Christ, M., Julliard, V., Pavirani, A., and Mehtali, M. (1997) Adenovirus-mediated gene transfer: influence of transgene, mouse strain and type of immune response on persistence of transgene expression. Gene Ther. 4, 473–482.

    Article  PubMed  CAS  Google Scholar 

  41. Barr, D., Tubb, J., Ferguson, D., Scaria, A., Lieber, A., Wilson, C., et al. (1995) Strain related variations in adenovirally mediated transgene expression from mouse hepatocytes in vivo: comparisons between immunocompetent and immunodeficient inbred strains. Gene Ther. 2, 151–155.

    PubMed  CAS  Google Scholar 

  42. Fields, P. A., Armstrong, E., Hagstrom, J. N., Arruda, V. R., Murphy, M. L., Farrell, J. P., et al. (2001) Intravenous administration of an E1/E3-deleted adenoviral vector induces tolerance to factor IX in C57BL/6 mice. Gene Ther. 8, 354–361.

    Article  PubMed  CAS  Google Scholar 

  43. Wen, X. Y., Bai, Y., and Stewart, A. K. (2001) Adenovirus-mediated human endostatin gene delivery demonstrates strain-specific antitumor activity and acute dose-dependent toxicity in mice. Hum. Gene Ther. 12, 347–358.

    Article  PubMed  CAS  Google Scholar 

  44. Smith, T., Idamaknati, N., Kylefjord, H., Rollence, M., King, L., Kaloss, M., et al. (2002) In vivo hepatic adenoviral gene delivery occurs in a CAR independent fashion. Mol. Ther. 5, 770–779.

    Article  PubMed  CAS  Google Scholar 

  45. Bristol, J. A., Shirley, P., Idamakanti, N., Kaleko, M., and Connelly, S. (2000) In vivo dose threshold effect of adenovirus-mediated factor VIII gene therapy in hemophiliac mice. Mol. Ther. 2, 223–232.

    Article  PubMed  CAS  Google Scholar 

  46. Connelly, S., Gardner, J. M., Lyons, R. M., McClelland, A., and Kaleko, M. (1996) Sustained expression of therapeutic levels of human factor VIII in mice. Blood 87, 4671–4677.

    PubMed  CAS  Google Scholar 

  47. Tao, N., Gao, G. P., Parr, M., Johnston, J., Baradet, T., Wilson, J. M., et al. (2001) Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver. Mol. Ther. 3, 28–35.

    Article  PubMed  CAS  Google Scholar 

  48. Mittereder, N., March, K. L., and Trapnell, B. C. (1996) Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J. Virol. 70, 7498–7509.

    PubMed  CAS  Google Scholar 

  49. Connelly, S., Smith, T. A., Dhir, G., Gardner, J. M., Mehaffey, M. G., Zaret, K. S., et al. (1995) In vivo gene delivery and expression of physiological levels of functional human factor VIII in mice. Hum. Gene Ther. 6, 185–193.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Advanced Vision Therapies Inc., Rockville, MD

    Sheila Connelly

  2. Genetic Therapy Inc., Gaithersburg, MD

    Christine Mech

Authors
  1. Sheila Connelly
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  2. Christine Mech
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Editors and Affiliations

  1. Bio-Rad Laboratories, Hercules, CA

    William C. Heiser

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© 2004 Humana Press Inc., Totowa, NJ

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Connelly, S., Mech, C. (2004). Delivery of Adenoviral DNA to Mouse Liver. In: Heiser, W.C. (eds) Gene Delivery to Mammalian Cells. Methods in Molecular Biology™, vol 246. Humana Press. https://doi.org/10.1385/1-59259-650-9:37

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  • DOI: https://doi.org/10.1385/1-59259-650-9:37

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-095-3

  • Online ISBN: 978-1-59259-650-8

  • eBook Packages: Springer Protocols

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