Radio-responsive recA promoter significantly increases TNFalpha production in recombinant clostridia after 2 Gy irradiation.
One of the major problems with gene therapy today is the lack of tumour specificity. The use of anaerobic apathogenic clostridia as a gene transfer system can target anoxic areas within the tumour. These bacteria can be genetically modified to express therapeutic proteins such as TNFalpha locally in the tumour. As shown in our results, ionising irradiation can be used in clostridia to activate genes encoding cytotoxic agents under control of a radiation-inducible promoter. A 44% significant increase (P < 0.05) in TNFalpha secretion was seen 3.5 h after a single dose of 2 Gy. A second dose of 2 Gy was also capable of repeating gene activation and gave a significant increase of TNFalpha production of 42% (P < 0.05). These results provide evidence that spatial and temporal control of gene expression can be achieved using a radio-inducible promoter. Repetitive gene activation was feasible with a second dose of 2 Gy, indicating that fractionated radiotherapy could lead to repeated gene induction resulting in prolonged and enhanced protein expression. Gene targeting by ionising radiation could thus provide a new means of increasing the therapeutic ratio in cancer treatment.
Nuyts
S
S
Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Belgium.
Van Mellaert
L
L
Theys
J
J
Landuyt
W
W
Bosmans
E
E
Anné
J
J
Lambin
P
P
D003013:Clostridium / Q000235:genetics* / Q000528:radiation effects; D019583:Dose Fractionation, Radiation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005786:Gene Expression Regulation / Q000528:radiation effects*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009369:Neoplasms / Q000532:radiotherapy / Q000628:therapy*; D011938:Rec A Recombinases / Q000235:genetics*; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000235:genetics*
D014409:Tumor Necrosis Factor-alpha; D011938:Rec A Recombinases