1. ¹Department of Urology, University of California at Los Angeles School of Medicine, Los Angeles, California 90095, USA
2. ²Department of Biological Chemistry, University of California at Los Angeles School of Medicine, Los Angeles, California 90095, USA
3. ³Crump Institute of Molecular Imaging and Department of Molecular and Medical Pharmacology, University of California at Los Angeles School of Medicine, Los Angeles, California 90095, USA

Correspondence: Lily Wu, Fax: (310) 206-5343. E-mail: LWu@mednet.ucla.edu

^*Present address: Department of Radiology and Bio-X Program, Stanford University, Stanford, CA.

Received 16 June 2003; Accepted 20 August 2003.

Abstract

Gene expression-based imaging coupled to gene therapy will permit the prediction of therapeutic outcome. A significant challenge for successful gene therapy is to achieve a high-level of specific gene expression; however, tissue-specific promoters are weak. We postulate that if the weak activity of tissue-specific promoters can be amplified to the levels of strong viral promoters, which have been successful in preclinical scenarios, while retaining specificity, the therapeutic index of gene therapy can be greatly augmented. With this in mind, we developed a two-step transcriptional activation (TSTA) system. In this two-tiered system, a modified prostate-specific antigen promoter was employed to drive a potent synthetic transcriptional activator, GAL4-VP2. This, in turn, activated the expression of a GAL4-dependent reporter or therapeutic gene. Here we demonstrate that recombinant adenoviral vectors (Ads) in which we have incorporated prostate-targeted TSTA expression cassettes retain cell specificity and androgen responsiveness in cell culture and in animal models, as measured by noninvasive optical bioluminescence imaging. We investigated the mechanism of TSTA in different adenoviral configurations. In one configuration, both the activator and the reporter components are inserted into a single Ad (AdTSTA-FL). The activity of AdTSTA-FL exceeds that of a cytomegalovirus promoter-driven vector (AdCMV-FL), while maintaining tissue specificity. When the activator and reporter components are placed in two separate Ads, androgen induction is more robust than for the single AdTSTA-FL. Based on these findings, we hope to refine the TSTA Ads further to improve the efficacy and safety of prostate cancer gene therapy.