Abstract
We have previously shown that a dual system for controlling gene expression that relies both on transcriptional regulation and DNA recombination mediated by the site-directed recombinase, Flp, effectively controls the expression of a gene encoding diphtheria toxin (DT-A). In this study, we investigated the use of a chimeric modified enhancer/promoter sequence of the human prostate-specific antigen (PSA) gene to regulate DT-A expression in human prostate cancer cells in culture, in xenografts derived from these cells, and in autochthonous tumors in TRAMP mice. Following adenoviral delivery of DNA encoding PSA promoter-driven Flp recombinase and DT-A, we demonstrate that this transcriptional/DNA recombination control strategy effectively activates DT-A expression in a manner that correlates with the amount of PSA and androgen in cells. Significantly, the size of xenografts was reduced by 50%, and tumor cells in TRAMP mice died following intratumoral injection of DT-A viruses. Direct injection of virally-delivered DT-A into normal mouse prostates resulted in a dramatic reduction in the size of the gland. Our results suggest that the PSA promoter-driven Flp recombinase regulatory system will allow for targeted death of PSA-expressing cells. When combined with newly developed strategies for targeted gene delivery, this approach holds promise as an effective systemically-administered therapy for metastatic prostate cancer.
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Acknowledgements
We thank Cheryl Hobbs for helpful comments on the manuscript, Lisa Laury-Kleintop for assistance with fluorescent microscopy, and Gwendolyn Gilliard for excellent technical assistance. We are grateful to Lily Wu (UCLA) for a plasmid containing the PSE-BC promoter, Michael Carey (UCLA) for pBCVP2G5-lucNSN, Susan Dymecki (Harvard) for pFRT2neolacZ, A Francis Stewart (EMBL, Heidelberg) for p22EDT1, J Miyazaki (Kyoto U) for pCX-EGFP, and R Rodriguez (Johns Hopkins U) for 293DTRP#2 cells. This work was supported by NIH Grant CA90841 (JAS).
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Peng, W., Chen, J., Huang, YH. et al. Tightly-regulated suicide gene expression kills PSA-expressing prostate tumor cells. Gene Ther 12, 1573–1580 (2005). https://doi.org/10.1038/sj.gt.3302580
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DOI: https://doi.org/10.1038/sj.gt.3302580
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