The expression of bacterial nitroreductase in eukaryotic cells results in their killing by the prodrug CB1954.
The enzyme nitroreductase, isolated from Escherichia coli B, converts CB1954 ((5-aziridin-1-yl)-2,4-dinitro-benzamide) into a cytotoxic DNA interstrand cross-linking agent. The E. coli B gene (nfnB, NTR) encoding nitroreductase (NTR) was cloned into eukaryotic expression vectors. When driven by a CMV promoter, 5-10% of the stably transfected mouse fibroblasts expressed the NTR enzyme. These cells were killed at a concentration of 20 microM CB1954 in comparison to nonexpressing cells which were killed at a much higher concentration (500 microM). We subsequently generated transgenic mice to test the prodrug system in vivo. Nitroreductase was expressed specifically in T cells driven by the control elements of the human CD2 locus. Upon CB1954 treatment, transgenic mice show extensive cell depletion in thymus and spleen (14-16% of normal cell numbers), whereas all other tissues are unaffected by prodrug administration. These results raise the possibility of using the NTR gene in anticancer therapy.
|Keywords||0 (Antineoplastic Agents), 0 (Aziridines), 0 (Prodrugs), 21919-05-1 (5-(1-aziridinyl)-2,4-dinitrobenzamide), Animals, Antineoplastic Agents/metabolism/*pharmacology, Apoptosis/drug effects, Aziridines/metabolism/*pharmacology, Cell Culture, EC 1.7.- (Nitroreductases), Escherichia coli/enzymology/genetics, Gene Expression, Gene Therapy/*methods, Mice, Mice, Transgenic, Nitroreductases/*genetics/metabolism, Prodrugs/metabolism/*pharmacology, Support, Non-U.S. Gov't, T-Lymphocytes/drug effects/enzymology, Transfection|
Drabek, D.D., Guy, J.A., & Grosveld, F.G.. (1997). The expression of bacterial nitroreductase in eukaryotic cells results in their killing by the prodrug CB1954.. Gene Therapy, 4, 93–100. Retrieved from http://hdl.handle.net/1765/2536