Optimization of adenoviral vectors to direct highly amplified prostate-specific expression for imaging and gene therapy.
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.
Department of Urology, University of California at Los Angeles School of Medicine, Los Angeles, California 90095, USA.
Makoto
M
Sato
Mai
M
Johnson
Liqun
L
Zhang
Baohui
B
Zhang
Kim
K
Le
Sanjiv S
SS
Gambhir
Michael
M
Carey
Lily
L
Wu
D000256:Adenoviridae / Q000235:genetics*; D000728:Androgens / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D051379:Mice; D016513:Mice, SCID; D018715:Microscopy, Video; D008957:Models, Genetic; D009368:Neoplasm Transplantation; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000628:therapy*; D013997:Time Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D000728:Androgens