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Replicating adenoviral vector–mediated transfer of a heat-inducible double suicide gene for gene therapy

Cancer Gene Therapy volume 8, pages 397–404 (2001)Cite this article

Abstract

Tumor cells that express a fusion gene of Escherichia coli cytosine deaminase (CD) and herpes simplex virus type 1 thymidine kinase (TK) sequences activate and are subsequently killed by the nontoxic prodrugs 5-fluorocytosine and ganciclovir. We have previously developed a recombinant adenovirus containing the CD-TK fusion gene controlled by the human inducible heat shock protein 70 promoter so that heat at 41°C for 1 hour induces therapeutic gene expression. This adenovirus effectively transduces heat-inducible expression of the CD-TK gene into human prostate carcinoma cells. However, because a limited number of cells in a tumor can actually be infected, we created a replicating adenoviral vector to increase CD-TK gene expression. This vector is a replication-competent, E1B-attenuated adenoviral vector containing the hsp70 promoter–driven CD-TK gene (Ad.E1A(+)HS-CDTK). When human prostate adenocarcinoma DU-145 cells (mutant p53) were infected with the virus at a multiplicity of infection (MOI) of 1 or 10, the viral replication was detected within 2 days at both MOIs. Similar results were observed in human colorectal carcinoma CX-1 cells. When DU-145 cells were infected with the virus at an MOI of 10, incubated for 24 hours, heated at 41°C for 4 hours, and then harvested 20 hours later, Western blot analysis demonstrated that this virus successfully produced viral E1A proteins and heat shock stimulated the CD-TK gene expression by 12.3-fold. In addition, Ad.E1A(+)HS-CDTK effectively suppressed cell proliferation by viral cytopathic effect). Unlike with a replication-incompetent virus (Ad.HS-CDTK), the cytopathic effect of the virus and cytotoxicity in the presence of the prodrugs were still observed even at low MOI (MOI=1.0). Cancer Gene Therapy (2001) 8, 397–404

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Acknowledgements

This work was supported by the following grants: NCI CA48000, CA44550, CA42857, CA44704, and Elsa U. Pardee Foundation.

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Authors and Affiliations

  1. Department of Pharmacology and Cancer Institute, University of Pittsburgh, Pittsburgh, 15213, Pennsylvania

    Yong J Lee & Paul Battle

  2. Department of Radiation Oncology, William Beaumont Hospital, Royal Oak, 48073, Michigan

    Sandra S Galoforo & Peter M Corry

  3. Department of Microbiology, Ulsan University, School of Medicine, Seoul, South Korea

    Heurian Lee

  4. Department of Surgery, University of Texas Health Science Center, San Antonio, 78284, Texas

    J Milburn Jessup

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  1. Yong J Lee
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  2. Sandra S Galoforo
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  3. Paul Battle
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  4. Heurian Lee
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  5. Peter M Corry
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  6. J Milburn Jessup
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Correspondence to Heurian Lee.

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Lee, Y., Galoforo, S., Battle, P. et al. Replicating adenoviral vector–mediated transfer of a heat-inducible double suicide gene for gene therapy. Cancer Gene Ther 8, 397–404 (2001). https://doi.org/10.1038/sj.cgt.7700310

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