Development of targeted therapy for bladder cancer mediated by a double promoter plasmid expressing diphtheria toxin under the control of IGF2-P3 and IGF2-P4 regulatory sequences

摘要

Background: The human IGF2-P3 and IGF2-P4 promoters are highly active in bladder carcinoma, while existing at a nearly undetectable level in the surrounding normal tissue. A double promoter DTA-expressing vector was created, carrying on a single construct two separate genes expressing diphtheria toxin A-fragment (DTA), from two different regulatory sequences, selected from the cancer-specific promoters IGF2-P3 and IGF2-P4. Methods: IGF2-P3 and IGF2-P4 expression was tested in samples of urothelial carcinoma (UC) of the bladder (n=67) by RT-PCR or by ISH. The therapeutic potential of single promoter expression vectors (P3-DTA and P4-DTA) was tested and compared to the double promoter toxin vector P4-DTA-P3-DTA in UC cell lines and in heterotopic and orthotopic animal models for bladder cancer. Results: Nearly 86% of UC patients highly expressed IGF2-P4 and IGF2-P3, as determined by ISH. The double promoter vector (P4-DTA-P3-DTA) exhibited superior ability to inhibit tumor development by 68% (P=0.004) compared to the single promoter expression vectors, in heterotopic bladder tumors. The average size of the P4-DTA-P3-DTA bladder tumors (in orthotopically treated mice) was 83% smaller (P<0.001) than that of the control group. Conclusions: Overall, the double promoter vector exhibited enhanced anti-cancer activity relative to single promoter expression vectors carrying either gene alone. Our findings show that bladder tumors may be successfully treated by intravesical instillation of the double promoter vector P4-DTA-P3-DTA.