Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant (original) (raw)

Cancer Gene Therapy volume 7, pages 66–73 (2000)Cite this article

Abstract

Tumor-directed gene therapy, such as “suicide gene” therapy, requires high levels of gene expression in a high percentage of tumor cells in vivo to be effective. Current vector strategies have been ineffective in achieving these goals. This report introduces the attenuated (thymidine kinase (TK)-negative) replication-competent vaccinia virus (VV) as a potential vector for tumor-directed gene therapy by studying the biodistribution of VV in animal tumor models. A TK-deleted recombinant VV (Western Reserve strain) expressing luciferase on a synthetic promoter was constructed. Luciferase activity was measured in vitro after transduction of a variety of human and murine tumor cell lines and in vivo after intraperitoneal (i.p.) delivery in C57BL/6 mice with 7-day i.p. tumors (106 MC-38 cells). Three other in vivo tumor models were examined for tumor-specific gene expression after intravenous delivery of VV (human melanoma in nude mice, adenocarcinoma liver metastasis in immunocompetent mice, and subcutaneous sarcoma in the rat). In addition, a replication-incompetent vaccinia (1 μg of psoralen and ultraviolet light, 365 nm, 4 minutes) was tested in vitro and in vivo and compared with active virus. Luciferase activity in i.p. tumors at 4 days after i.p. injection of VV was >7000-fold higher than lung, >3000-fold higher than liver, and >250-fold higher than ovary. In addition, intravenous injection of VV resulted in markedly higher tumor luciferase activity compared with any other organ in every model tested (up to 188,000-fold higher than liver and 77,000-fold higher than lung). Inactivation of the virus resulted in negligible gene expression in vivo. In summary, VV has a high transduction efficiency in tumor cells with high levels of gene expression. The results suggest a selective in vivo replication of TK-deleted VV in tumor cells. Replication competent, TK-deleted VV appears to be an ideal vector for testing the in vivo delivery of toxic genes to tumor cells.

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Authors and Affiliations

  1. Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, 20892, Maryland
    Markus Puhlmann, Charles K Brown, Michael Gnant, James Huang, Steven K Libutti, H Richard Alexander & David L Bartlett

Authors

  1. Markus Puhlmann
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  2. Charles K Brown
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  3. Michael Gnant
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  4. James Huang
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  5. Steven K Libutti
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  6. H Richard Alexander
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  7. David L Bartlett
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Corresponding author

Correspondence toDavid L Bartlett.

Additional information

This paper was presented in part at the 88th Annual Meeting of the American Association for Cancer Research, March 1997, San Diego, California.

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Puhlmann, M., Brown, C., Gnant, M. et al. Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant.Cancer Gene Ther 7, 66–73 (2000). https://doi.org/10.1038/sj.cgt.7700075

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