title: Telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells to the pro-drug CB1954.
abstract: Telomerase activation is considered to be a critical step in cancer progression due to its role in cellular immortalization. The prevalence of telomerase expression in human cancers makes it an attractive candidate for new mechanism-based targets for cancer therapy. The selective killing of cancer cells can be achieved by gene-directed enzyme pro-drug therapy (GDEPT). In this study we have tested the feasibility of using the transcriptional regulatory 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 compared 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 lines exhibiting high levels of promoter activity. The in vivo relevance of these data was tested using two xenograft models (C33a and GLC4 cells). Significant tumour reduction was seen with both telomerase promoters and the promoter-specific patterns of sensitization observed in tissue culture were retained in xenograft models. Thus, telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells to the pro-drug CB1954.
authors.affiliation: CRC Department of Medical Oncology, University of Glasgow, CRC Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, UK.
authors.forename: J A
authors.initials: JA
authors.lastname: Plumb
authors.forename: A
authors.initials: A
authors.lastname: Bilsland
authors.forename: R
authors.initials: R
authors.lastname: Kakani
authors.forename: J
authors.initials: J
authors.lastname: Zhao
authors.forename: R M
authors.initials: RM
authors.lastname: Glasspool
authors.forename: R J
authors.initials: RJ
authors.lastname: Knox
authors.forename: T R
authors.initials: TR
authors.lastname: Evans
authors.forename: W N
authors.initials: WN
authors.lastname: Keith
mesh_terms: D000818:Animals; D001388:Aziridines / Q000494:pharmacology* / Q000627:therapeutic use; D001419:Bacteria / Q000201:enzymology; D018288:Carcinoma, Small Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D009601:Nitroreductases / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use*; D011355:Prodrugs / Q000494:pharmacology* / Q000627:therapeutic use; D011401:Promoter Regions, Genetic / Q000235:genetics; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes / Q000235:genetics; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy
chemical_list: D001388:Aziridines; D011355:Prodrugs; C100099:tretazicar; D009601:Nitroreductases; D019098:Telomerase