Skip to main content
SpringerLink
Log in

Construction of Targeted and Armed Oncolytic Adenoviruses

  • Protocol
  • First Online:
Oncolytic Viruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 797))

Abstract

Oncolytic (replication-competent) adenoviruses (Ads) represent the most advanced platform for cancer gene therapy. These viral vectors ablate tumors by killing tumor cells in the process of virus replication. As progeny virions are released, they infect remaining cancer cells, generating a bystander effect. Ads engineered for increased cancer specificity produce less damage to normal tissues. First-generation oncolytic Ads have demonstrated acceptable levels of safety while the efficacy was observed only in combination with chemotherapy and/or radiation. Second-generation oncolytic Ads are armed with therapeutic transgenes to increase release, spread, and bystander effect for enhancing the efficacy. Third-generation oncolytic Ads are armed vectors with capsid modifications for transductional detargeting from normal tissues and targeting to cancer cells. Chemical modification of the capsid additionally improves therapeutic window. Here, we describe methods for generation and characterization of advanced-generation oncolytic Ads.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Graham, F. L., Smiley, J., Russell, W. C., and Nairn, R. (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 36, 59–74.

    Article  CAS  Google Scholar 

  2. Huebner, R. J., Rowe, W. P., Schatten, W. E., Smith, R. R., and Thomas, L. B. (1956) Studies on the use of viruses in the treatment of carcinoma of the cervix. Cancer. 9, 1211–1218.

    Article  CAS  Google Scholar 

  3. Liu, T. C., Galanis, E., and Kirn, D. (2007) Clinical trial results with oncolytic virotherapy: a century of promise, a decade of progress. Nat Clin Pract Oncol. 4, 101–117.

    Article  CAS  Google Scholar 

  4. Yamamoto, M. and Curiel, D. T. (2010) Current issues and future directions of oncolytic adenoviruses. Mol Ther. 18, 243–250.

    Article  CAS  Google Scholar 

  5. Doronin, K., Toth, K., Kuppuswamy, M., Ward, P., Tollefson, A. E., and Wold, W. S. (2000) Tumor-specific, replication-competent adenovirus vectors overexpressing the adenovirus death protein. J Virol. 74, 6147–6155.

    Article  CAS  Google Scholar 

  6. Doronin, K., Kuppuswamy, M., Toth, K., Tollefson, A. E., Krajcsi, P., Krougliak, V., et al. (2001) Tissue-specific, tumor-selective, replication-competent adenovirus vector for cancer gene therapy. J Virol. 75, 3314–3324.

    Article  CAS  Google Scholar 

  7. Shashkova, E. V., Kuppuswamy, M. N., Wold, W. S., and Doronin, K. (2008) Anticancer activity of oncolytic adenovirus vector armed with IFN-alpha and ADP is enhanced by pharmacologically controlled expression of TRAIL. Cancer Gene Ther. 15, 61–72.

    Article  CAS  Google Scholar 

  8. Shayakhmetov, D. M., Papayannopoulou, T., Stamatoyannopoulos, G., and Lieber, A. (2000) Efficient gene transfer into human CD34(+) cells by a retargeted adenovirus vector. J Virol. 74, 2567–2583.

    Article  CAS  Google Scholar 

  9. Doronin, K., Shashkova, E. V., May, S. M., Hofherr, S. E., and Barry, M. A. (2009) Chemical modification with high molecular weight polyethylene glycol reduces transduction of hepatocytes and increases efficacy of intravenously delivered oncolytic adenovirus. Hum Gene Ther. 20, 975–988.

    Article  CAS  Google Scholar 

  10. Shashkova, E. V., Spencer, J. F., Wold, W. S., and Doronin, K. (2007) Targeting interferon-alpha increases antitumor efficacy and reduces hepatotoxicity of E1A-mutated spread-enhanced oncolytic adenovirus. Mol Ther. 15, 598–607.

    Article  CAS  Google Scholar 

  11. Sauthoff, H., Pipiya, T., Heitner, S., Chen, S., Bleck, B., Reibman, J., et al. (2004) Impact of E1a modifications on tumor-selective adenoviral replication and toxicity. Mol Ther. 10, 749–757.

    Article  CAS  Google Scholar 

  12. Krougliak, V. and Graham, F. L. (1995) Development of cell lines capable of complementing E1, E4, and protein IX defective adenovirus type 5 mutants. Hum Gene Ther. 6, 1575–1586.

    Article  CAS  Google Scholar 

  13. Chen, C. and Okayama, H. (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 7, 2745–2752.

    Article  CAS  Google Scholar 

  14. Kuppuswamy, M., Spencer, J. F., Doronin, K., Tollefson, A. E., Wold, W. S., and Toth, K. (2005) Oncolytic adenovirus that overproduces ADP and replicates selectively in tumors due to hTERT promoter-regulated E4 gene expression. Gene Ther. 12, 1608–1617.

    Article  CAS  Google Scholar 

  15. Bett, A. J., Haddara, W., Prevec, L., and Graham, F. L. (1994) An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3. Proc Natl Acad Sci USA. 91, 8802–8806.

    Article  CAS  Google Scholar 

  16. Gaggar, A., Shayakhmetov, D. M., and Lieber, A. (2003) CD46 is a cellular receptor for group B adenoviruses. Nat Med. 9, 1408–1412.

    Article  CAS  Google Scholar 

  17. Sova, P., Ren, X. W., Ni, S., Bernt, K. M., Mi, J., Kiviat, N., et al. (2004) A tumor-targeted and conditionally replicating oncolytic adenovirus vector expressing TRAIL for treatment of liver metastases. Mol Ther. 9, 496–509.

    Article  CAS  Google Scholar 

  18. Mizuguchi, H. and Kay, M. A. (1998) Efficient construction of a recombinant adenovirus vector by an improved in vitro ligation method. Hum Gene Ther. 9, 2577–2583.

    Article  CAS  Google Scholar 

  19. Kalyuzhniy, O., Di Paolo, N. C., Silvestry, M., Hofherr, S. E., Barry, M. A., Stewart, P. L., et al. (2008) Adenovirus serotype 5 hexon is critical for virus infection of hepatocytes in vivo. Proc Natl Acad Sci USA. 105, 5483–5488.

    Google Scholar 

  20. Alba, R., Bradshaw, A. C., Parker, A. L., Bhella, D., Waddington, S. N., Nicklin, S. A., et al. (2009) Identification of coagulation factor (F)X binding sites on the adenovirus serotype 5 hexon: effect of mutagenesis on FX interactions and gene transfer. Blood. 114, 965–971.

    Article  CAS  Google Scholar 

  21. Shashkova, E. V., May, S. M., Doronin, K., and Barry, M. A. (2009) Expanded anticancer therapeutic window of hexon-modified oncolytic adenovirus. Mol Ther. 17, 2121–2130.

    Article  CAS  Google Scholar 

  22. Campos, S. K. and Barry, M. A. (2004) Rapid construction of capsid-modified adenoviral vectors through bacteriophage lambda Red recombination. Hum Gene Ther. 15, 1125–1130.

    Article  CAS  Google Scholar 

  23. Graham, F. L. (1987) Growth of 293 cells in suspension culture. J Gen Virol. 68 (Pt 3), 937–940.

    Article  Google Scholar 

  24. O’Riordan, C. R., Lachapelle, A., Delgado, C., Parkes, V., Wadsworth, S. C., Smith, A. E., et al. (1999) PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. Hum Gene Ther. 10, 1349–1358.

    Article  Google Scholar 

  25. Shashkova, E. V., Doronin, K., Senac, J. S., and Barry, M. A. (2008) Macrophage depletion combined with anticoagulant therapy increases therapeutic window of systemic treatment with oncolytic adenovirus. Cancer Res. 68, 5896–5904.

    Article  CAS  Google Scholar 

  26. Duncan, S. J., Gordon, F. C., Gregory, D. W., McPhie, J. L., Postlethwaite, R., White, R., et al. (1978) Infection of mouse liver by human adenovirus type 5. J Gen Virol. 40, 45–61.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported by R01CA141439 to D.M. Shayakhmetov.

Author information

Authors and Affiliations

  1. Division of Medical Genetics, University of Washington, Seattle, WA, USA

    Konstantin Doronin

  2. University of Washington, Seattle, WA, USA

    Dmitry M. Shayakhmetov

Authors
  1. Konstantin Doronin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dmitry M. Shayakhmetov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Konstantin Doronin .

Editor information

Editors and Affiliations

  1. Jennerex Biotherapeutics, Inc., Market Street 1, San Francisco, 94105, California, USA

    David H. Kirn

  2. School of Medicine, Dept. Pathology & Immunology, Washington University, S. Euclid Ave. 660, St.Louis, 63110, Missouri, USA

    Ta-Chiang Liu

  3. Hillman Cancer Center, University of Pittsburgh, Centre Avenue 5117, Pittsburgh, 15213, Pennsylvania, USA

    Stephen H. Thorne

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Doronin, K., Shayakhmetov, D.M. (2012). Construction of Targeted and Armed Oncolytic Adenoviruses. In: Kirn, D., Liu, TC., Thorne, S. (eds) Oncolytic Viruses. Methods in Molecular Biology, vol 797. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-340-0_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-340-0_3

  • Published:

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-1-61779-339-4

  • Online ISBN: 978-1-61779-340-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation