Telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells to the pro-drug CB1954

Telomerase activation is considered to be a critical step in cancer progres sion due to its role in cellular immortalization. The prevalence of telomer ase expression in human cancers makes it an attractive candidate for new me chanism-based targets for cancer therapy. The selective killing of cancer c ells can be achieved by gene-directed enzyme pro-drug therapy (GDEPT). In t his study we have tested the feasibility of using the transcriptional regul atory sequences from the hTERT and hTR genes to regulate expression of the bacterial nitroreductase enzyme in combination with the pro-drug CB1954 in a suicide gene therapy strategy. hTERT and hTR promoter activity was compar ed in a panel of 10 cell lines and showed a wide distribution in activity; low activity was observed in normal cells and telomerase-negative immortal ALT cell lines, with up to 300-fold higher activity observed in telomerase positive cancer lines. Placing the nitroreductase gene under the control of the telomerase gene promoters sensitized cancer cells in tissue culture to the pro-drug CB1954 and promoter activity was predictive of sensitization to the pro-drug (2-20-fold sensitization), with cell death restricted to li nes exhibiting high levels of promoter activity. The in vivo relevance of t hese data was tested using two xenograft models (C33a and GLC4 cells). Sign ificant tumour reduction was seen with both telomerase promoters and the pr omoter-specific patterns of sensitization observed in tissue culture were r etained in xenograft models. Thus, telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells t o the pro-drug CB1954.