Appropriate subcellular localisation of prodrug-activating enzymes has important consequences for suicide gene therapy.
Escherichia coli B nitroreductase (NR) has been expressed stably in MDA-MB-361 human breast adenocarcinoma cells either as the wild-type protein (wtNR), which is distributed evenly between the cytoplasmic and nuclear compartments, or targeted to the mitochondrion (mtNR). Whereas bacterial NR is active as a dimer, a proportion of wtNR is monomeric. In contrast, mtNR is mostly dimeric, suggesting that it adopts a more stable, native conformation. Despite this, when tested in gene-directed enzyme prodrug therapy cell cytotoxicity studies, cells expressing wtNR or mtNR had similar sensitivity to the prodrug CB1954 and mounted similar bystander killing effects. Furthermore, when short prodrug exposures were given, wtNR was more efficient at killing cells than mtNR. These data demonstrate that the site of enzyme expression and prodrug activation is an important variable that requires consideration in suicide gene therapy approaches.
Spooner
R A
RA
CRC Centre for Cancer Therapeutics, Institute of Cancer Research, Sutton, United Kingdom.
Maycroft
K A
KA
Paterson
H
H
Friedlos
F
F
Springer
C J
CJ
Marais
R
R
D000230:Adenocarcinoma; D000970:Antineoplastic Agents / Q000493:pharmacokinetics* / Q000633:toxicity; D001388:Aziridines / Q000493:pharmacokinetics* / Q000633:toxicity; D001483:Base Sequence; D001943:Breast Neoplasms; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D019281:Dimerization; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D008969:Molecular Sequence Data; D009601:Nitroreductases / Q000235:genetics / Q000378:metabolism*; D011355:Prodrugs / Q000493:pharmacokinetics* / Q000633:toxicity; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D001388:Aziridines; D011355:Prodrugs; D011994:Recombinant Proteins; C100099:tretazicar; D009601:Nitroreductases