Abstract
Virus-directed enzyme prodrug therapy (VDEPT) utilising cytosine deaminase (CD) converts 5-fluorocytosine (5-FC) into the chemotherapy agent, 5-fluorouracil (5-FU), and has entered into a clinical trial for metastatic colon cancer. To improve this system, a replication-deficient adenovirus, containing a bifunctional fusion gene, CD:uracil phosphoribosyltransferase (UPRT), was constructed (AdCDUPRT). UPRT enhances the conversion of 5-FU into its active metabolites, which inhibit DNA and RNA synthesis. In vitro, AdCDUPRT infection of colon cancer cells resulted in a marked increase in sensitisation to 5-FU, compared with AdCD-infected or uninfected cells. The corollary is a ∼100-fold and ∼10 000-fold increase in sensitisation to 5-FC in AdCDUPRT-infected cells, compared to AdCD-infected and uninfected cells, respectively. There was a strong bystander effect in vitro, 70% of tumour cells were killed by 5-FC when only 10% of cells expressed CDUPRT. In vivo, athymic mice with colon cancer xenografts treated with intratumoral AdCDUPRT and intraperitoneal 5-FC, significantly reduced tumour growth rates compared with untreated controls (P = 0.02), whereas AdCD/5-FC treated mice did not. At higher AdCDUPRT virus doses, 5-FC and 5-FU were equally effective at delaying tumour growth compared with controls. In summary, VDEPT for colon cancer utilising AdCDUPRT is more effective than AdCD and the bifunctional CDUPRT gene enables the use of either 5-FC or 5-FU as prodrugs.
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References
Ballantyne GH, Quin J . Surgical treatment of liver metastases in patients with colorectal cancer Cancer 1993 71: 4252–4266
Douillard JY et al. Irinotecan combined with fluorouracil compared with fluorouracil alone as first-line treatment for metastatic colorectal cancer: a multicentre randomised trial (published erratum appears in Lancet 2000 Apr 15;355(9212):1372) Lancet 2000 355: 1041–1047
Mullen CA, Kilstrup M, Blaese RM . Transfer of the bacterial gene for cytosine deaminase to mammalian cells confers lethal sensitivity to 5-fluorocytosine: a negative selection system Proc Nat Acad Sci USA 1992 89: 33–37
Austin EA, Huber BE . A first step in the development of gene therapy for colorectal carcinoma: cloning, sequencing, and expression of Escherichia coli cytosine deaminase Mol Pharm 1993 43: 380–387
Hirschowitz EA et al. In vivo adenovirus-mediated gene transfer of the Escherichia coli cytosine deaminase gene to human colon carcinoma-derived tumors induces chemosensitivity to 5-fluorocytosine Hum Gene Ther 1995 6: 1055–1063
Crystal RG et al. Phase I study of direct administration of a replication deficient adenovirus vector containing the E. coli cytosine deaminase gene to metastatic colon carcinoma of the liver in association with the oral administration of the pro-drug 5-fluorocytosine Hum Gene Ther 1997 8: 985–1001
Tiraby M et al. Concomitant expression of E. coli cytosine deaminase and uracil phosphoribosyltransferase improves the cytotoxicity of 5-fluorocytosine FEMS Microbiol Lett 1998 167: 41–49
Kanai F et al. Adenovirus-mediated transduction of Escherichia coli uracil phosphoribosyltransferase gene sensitizes cancer cells to low concentrations of 5-fluorouracil Cancer Res 1998 58: 1946–1951
Lan KH et al. Tumor-specific gene expression in carcinoembryonic antigen-producing gastric cancer cells using adenovirus vectors Gastroenterology 1996 111: 1241–1251
Recommended 5-fluorocytosine dose in patients British National Formulary 1999 37: 276–277
Pierrefite-Carle V et al. Cytosine deaminase/5-fluorocytosine-based vaccination against liver tumors: evidence of distant bystander effect J Nat Cancer Inst 1999 91: 2014–2019
Khuri FR et al. A controlled trial of intratumoral ONYX-015, a selectively-replicating adenovirus, in combination with cisplatin and 5-fluorouracil in patients with recurrent head and neck cancer Nat Med 2000 6: 879–885
Green NK et al. Sensitization of colorectal and pancreatic cancer cell lines to the prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) by retroviral transduction and expression of the E. coli nitroreductase gene Cancer Gene Ther 1997 4: 229–238
Latham JP, Searle PF, Mautner V, James ND . Prostate-specific antigen promoter/enhancer driven gene therapy for prostate cancer: construction and testing of a tissue-specific adenovirus vector Cancer Res 2000 60: 334–341
Fallaux FJ et al. Characterization of 911: a new helper cell line for the titration and propagation of early region 1-deleted adenoviral vectors Hum Gene Ther 1996 7: 215–222
Laird NM, Ware JH . Random-effects models for longitudinal data Biometrics 1982 38: 963–974
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This work was supported by the Cancer Research Campaign, UK and the Medical Research Council, UK.
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Chung-Faye, G., Chen, M., Green, N. et al. In vivo gene therapy for colon cancer using adenovirus-mediated, transfer of the fusion gene cytosine deaminase and uracil phosphoribosyltransferase. Gene Ther 8, 1547–1554 (2001). https://doi.org/10.1038/sj.gt.3301557
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DOI: https://doi.org/10.1038/sj.gt.3301557
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