Adenovirus-mediated suicide gene therapy for experimental bladder cancer

Mark A. Sutton, Scott A. Berkman, Shu-Hsia Chen, Andreas Block, Truong D. Dang, Michael W. Kattan, Thomas M. Wheeler, David R. Rowley, Savio L.C. Woo, Seth P. Lerner

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

Objectives. To determine the feasibility, safety, and efficacy of suicide gene therapy using adenoviral-mediated herpes simplex virus thymidine kinase gene (HSV-tk) and the prodrug ganciclovir (GCV) in a murine model of human transitional cell carcinoma. Methods. We used a replication-defective adenoviral construct containing the beta-galactosidase gene (ADV/Rous sarcoma virus [RSV]-beta-gal] as a control or ADV/RSV-tk as the therapeutic vector under the transcriptional control of the RSV long-terminal repeat promoter. Transduction efficiency was assessed in vitro by infection of MBT-2 cells with ADV/RSV-beta-gal at various multiplicities of infection (MOI) utilizing 5-bromo-4-chlor-3-indolyl-beta-D-galactoside (X-gal) staining. Sensitivity of MBT-2 cells to the therapeutic vector was determined after infection with ADV/RSV-tk with or without GCV. Subcutaneous tumors were established in syngeneic CSH/He female mice with 5 x 105 MBT-2 cells. Optimal dosing of ADV/RSV-tk was determined by direct percutaneous tumor injection with increasing viral doses and treatment with GCV. Treatment efficacy, long-term survival, and toxicity were determined in separate, similar, controlled experiments. Results. In vitro studies indicated greater than 95% transduction 96 hours after inoculation at an MOI of 3000 and a greater than 95% cell death rate with RSV-tk + GCV at an MOI of 61 or greater. In vivo experiments demonstrated an optimal viral dose of 3 x 108 plaque-forming units (pfu) and a greater than fourfold reduction in tumor growth for the animals treated with ADV/RSV-tk compared with control animals (P = 0.0013). Toxicity was limited to histologic evidence of hepatitis with ADV/RSV-tk doses greater than 3 x 108 pfu + GCV. Long-term survival of treatment animals was significantly increased over that of control animals (59%, P = 0.0001). Conclusions. ADV/RSV-tk with GCV treatment results in efficient gene transfer in vitro and provides effective therapy in experimental murine bladder cancer by significantly inhibiting tumor growth and improving host survival.

Original languageEnglish (US)
Pages (from-to)173-180
Number of pages8
JournalUrology
Volume49
Issue number2
DOIs
StatePublished - Feb 1 1997

ASJC Scopus subject areas

  • Urology

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