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Radio-responsive recA promoter significantly increases TNFα production in recombinant clostridia after 2 Gy irradiation

Gene Therapy volume 8, pages 1197–1201 (2001)Cite this article

Abstract

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 TNFα 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 TNFα 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 TNFα 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.

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References

  1. Weichselbaum RR, Hallahan DE, Sukhatme VP, Kufe DW . Gene therapy targeted by ionizing irradiation Int J Radiat Oncol Biol Phys 1992 24: 565–567

    Article  CAS  PubMed  Google Scholar 

  2. Weichselbaum RR et al. Gene therapy targeted by radiation preferentially radiosensitizes tumor cells Cancer Res 1994 54: 4266–4269

    CAS  PubMed  Google Scholar 

  3. Hallahan DE et al. Spatial and temporal control of gene therapy using ionizing irradiation Nate Med 1995 1: 786–791

    Article  CAS  Google Scholar 

  4. Joki J, Nakamura M, Ohno T . Activation of the radiosensitive EGR-1 promoter induces expression of the herpes simplex virus thymidine kinase gene and sensitivity of human glioma cells to ganciclovir Hum Gene Ther 1995 6: 1507–1513

    Article  CAS  PubMed  Google Scholar 

  5. Seung LP et al. Genetic radiotherapy overcomes tumor resistance to cytotoxic agents Cancer Res 1995 55: 5561–5565

    CAS  PubMed  Google Scholar 

  6. Takahashi T, Namiki Y, Ohno T . Induction of the suicide HSV TK gene by activation of the Egr 1 promoter with radioisotopes Hum Gene Ther 1997 8: 827–833

    Article  CAS  PubMed  Google Scholar 

  7. Manome Y et al. Transgene expression in malignant glioma using a replication-defective adenoviral vector containing the Egr-1 promoter: activation by ionizing irradiation or uptake of radioactive iododeoxyridine Hum Gene Ther 1998 9: 1409–1417

    Article  CAS  PubMed  Google Scholar 

  8. Kawashita Y et al. Regression of hepatocellular carcinoma in vitro and in vivo by radiosensitizing suicide gene therapy under the inducible and spatial control of radiation Hum Gene Ther 1999 10: 1509–1519

    Article  CAS  PubMed  Google Scholar 

  9. Marples B et al. Development of synthetic promoters for radiation-mediated gene therapy Gene Therapy 2000 7: 511–517

    Article  CAS  PubMed  Google Scholar 

  10. Larrick JW, Wright SC . Cytotoxic mechanism of tumour necrosis factor-α FASEB J 1990 4: 3215–3223

    Article  CAS  PubMed  Google Scholar 

  11. Old LJ . Tumour necrosis factor Science 1985 230: 630–636

    Article  CAS  PubMed  Google Scholar 

  12. Staba MJ et al. Adenoviral TNF-α gene therapy and radiation damage tumor vasculature in a human malignant glioma xenograft Gene Therapy 1998 5: 293–300

    Article  CAS  PubMed  Google Scholar 

  13. Hallahan DE, Beckett MA, Kufe D, Weichselbaum RR . The interaction between recombinant human tumor necrosis factor and radiation in 13 human tumor cell lines Int J Radiat Oncol Biol Phys 1990 19: 69–74

    Article  CAS  PubMed  Google Scholar 

  14. Lambin P et al. Colonisation of Clostridium in the body is restricted to hypoxic and necrotic areas of tumours Anaerobe 1998 4: 183–188

    Article  CAS  PubMed  Google Scholar 

  15. Theys J et al. Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha Appl Environ Microbiol 1999 65: 4295–4300

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Theys J et al. Specific targeting of cytosine deaminase to solid tumors by engineered Clostridium acetobutylicum Cancer Gene Ther (in press)

  17. Fox ME et al. Anaerobic bacteria as delivery system for cancer gene therapy: in vitro activation of 5-fluorocytosine by genetically engineered clostridia Gene Therapy 1996 3: 173–178

    CAS  PubMed  Google Scholar 

  18. Lemmon MJ et al. Anaerobic bacteria as a gene delivery system that is controlled by the tumor microenvironment Gene Therapy 1997 4: 791–796

    Article  CAS  PubMed  Google Scholar 

  19. Nuyts S et al. Increasing specificity of anti-tumor therapy: cytotoxic protein delivery by non-pathogenic clostridia under regulation of radio-induced promoters Anticancer Res 2001 21: 857–862

    CAS  PubMed  Google Scholar 

  20. Nuyts S et al. The use of radio-induced bacterial promoters in anaerobic conditions: a means to control gene expression in Clostridium-mediated therapy for cancer Radiat Res 2001 155: 716–723

    Article  CAS  PubMed  Google Scholar 

  21. Nuyts S et al. Manipulation radio-inducibility of recA promoter in Clostridium Appl Environ Microbiol (submitted)

  22. Zimmerman RJ, Chan A, Leadon SA . Oxidative damage in murine tumor cells treated in vitro by recombinant human TNF Cancer Res 1989 49: 1644–1648

    CAS  PubMed  Google Scholar 

  23. Theys J et al. Improvement of Clostridium tumour targeting vectors evaluated in rat rhabdomyosarcomas FEMS Immunol Med Microbiol 2001 30: 37–41

    Article  CAS  PubMed  Google Scholar 

  24. Platt J et al. Antitumour effects of geneticaly engineered Salmonella in combination with radiation Eur J Cancer 2000 36: 2397–2402

    Article  CAS  PubMed  Google Scholar 

  25. Pawelek JK, Low KB, Bermudes D . Tumor-targeted Salmonella as a novel anticancer vector Cancer Res 1997 57: 4537–4544

    CAS  PubMed  Google Scholar 

  26. Yazawa K et al. Bifidobacterium longum as a delivery system for cancer gene therapy: selective localization and growth in hypoxic tumors Cancer Gene Ther 2000 7: 269–274

    Article  CAS  PubMed  Google Scholar 

  27. Oultram J et al. Introduction of plasmids into whole cells of Clostridium acetobutylicum by electroporation FEMS Microbiol Lett 1988 56: 83–88

    Article  CAS  Google Scholar 

  28. Sambrook JE, Fritsch EF, Maniatis T . Molecular Cloning: a Laboratory Manual, 2nd edn Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York 1989

    Google Scholar 

  29. Mermelstein LD, Papoutsakis ET . In vivo methylation in Escherichia coli by the Bacillus subtillus phage Φ3T I methyltransferase to protect plasmids from restriction upon transformation of Clostridium acetobutylicum ATCC824 Appl Environ Microbiol 1993 59: 1077–1081

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Mermelstein LD, Welker NE, Bennett GE, Papoutsakis ET . Expression of cloned homologous fermentative genes in Clostridium acetobutylicum ATCC824 BioTechnology 1992 10: 190–195

    CAS  PubMed  Google Scholar 

  31. Nakotte S, Schaffer M, Böhringer M, Dürre P . Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM792 Appl Microbiol Biotech 1998 50: 564–567

    Article  CAS  Google Scholar 

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Acknowledgements

We acknowledge the financial support from ‘Het Fonds voor Wetenschappelijk Onderzoek-Vlaanderen’, ‘Verkennende Internationale Samenwerking’ and ‘Het KU Leuven Onderzoeksfonds’. Sandra Nuyts is research fellow of ‘IWT’ (Vlaams Instituut voor de bevordering van het Wetenschappelijk-Technologisch onderzoek in de industrie). We wish to thank Raf Berghmans, DiaMed EuroGen, Tessenderlo, Belgium for providing the ELISA kits.

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Authors and Affiliations

  1. Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Leuven, Belgium

    S Nuyts, L Van Mellaert, J Theys & J Anné

  2. Laboratory of Experimental Radiobiology, University Hospital Gasthuisberg, Leuven, Belgium

    S Nuyts & W Landuyt

  3. DiaMed EuroGen, Tessenderlo, Belgium

    E Bosmans

  4. Department of Radiation Oncology, RTIL, Academic Hospital Maastricht, University Maastricht, Maastricht, The Netherlands

    P Lambin

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  1. S Nuyts
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Nuyts, S., Van Mellaert, L., Theys, J. et al. Radio-responsive recA promoter significantly increases TNFα production in recombinant clostridia after 2 Gy irradiation. Gene Ther 8, 1197–1201 (2001). https://doi.org/10.1038/sj.gt.3301499

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  • DOI: https://doi.org/10.1038/sj.gt.3301499

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