Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Genetic prodrug activation therapy (GPAT) in two rat prostate models generates an immune bystander effect and can be monitored by magnetic resonance techniques

Gene Therapy volume 8, pages 557–567 (2001)Cite this article

Abstract

Treatment of hormone refractory prostate cancer requires new treatment strategies. Genetic prodrug activation therapy (GPAT) may provide a new therapeutic avenue. In this study the antitumour efficacy of the gene encoding herpes simplex virus thymidine kinase (HSVtk) activating the prodrug ganciclovir (GCV) was compared in two models of ectopic (subcutaneous) rat prostate cancer. Both models, which differ in their characteristics, were previously shown to be weakly immunogenic but susceptible to immunotherapy. Tumour cell lines were stably transfected with HSVtk and were rendered highly sensitive to GCV. Little or no bystander killing effect was observed by tk-transfected cells on wild-type cells in vitro. However, a significant in vivo bystander effect was observed suggesting an immune-mediated response. Indeed, such an immune response was capable of slowing the growth of distant wild-type tumours and increased overall animal survival. A T helper 1 immune response was generated as a result of GCV activation and cell kill, demonstrated by the secretion of IFNγ by cultured splenocytes in response to tumour cells. BrDU staining of tk-transfected cells treated with GCV in vitro suggested apoptotic cell death, but Annexin V staining was less marked for one of the cell lines. Serial in vivo monitoring by non-invasive magnetic resonance spectroscopy (MRS) of the tk-transfected MATLyLu tumours demonstrated a decreased ATP/Pi ratio (a measure of cell energy status) during growth and an increase in the ATP/Pi ratio during regression initiated by treatment with GCV. Further, significant differences were found in the phosphomonester (PME) to total phosphate (ΣP) ratios in treated compared with untreated tumours, a result rarely seen in animal models, but commonly observed in patients. This study showed that a Th1-biased immune response generated by killing prostate tumour cells with tk/GCV can kill distant as well as local wild-type tumour cells. These findings suggest that GPAT may have a potential application in patients with both confined and metastatic prostate cancer and MRS may provide a method of monitoring response to treatment.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Hrouda D, Perry M, Dalgleish AG . Gene therapy for prostate cancer Semin Oncol 1999 26: 455–471

    CAS  PubMed  Google Scholar 

  2. Pardoll DM . Cancer vaccines Nat Med 1998 4: 525–531

    Article  CAS  PubMed  Google Scholar 

  3. Simons JW et al. Induction of immunity to prostate cancer antigens: results of a clinical trial of vaccination with irradiated autologous prostate tumor cells engineered to secrete granulocyte–macrophage colony-stimulating factor using ex vivo gene transfer Cancer Res 1999 59: 5160–5168

    CAS  PubMed  Google Scholar 

  4. Tjoa BA et al. Follow-up evaluation of prostate cancer patients infused with autologous dendritic cells pulsed with PSMA peptides Prostate 1997 32: 272–278

    Article  CAS  PubMed  Google Scholar 

  5. Vesalainen S, Lipponen P, Talja M, Syrjanen K . Histological grade, perineural infiltration, tumour-infiltrating lymphocytes and apoptosis as determinants oflong-term prognosis in prostatic adenocarcinoma Eur J Cancer 1994 30A: 1797–1803

    Article  CAS  PubMed  Google Scholar 

  6. Hrouda D et al. Immunotherapy of advanced prostate cancer: a phase I/II trial using Mycobacterium vaccae (SRL172) Br J Urol 1998 82: 568–573

    Article  CAS  PubMed  Google Scholar 

  7. Maraveyas A et al. Possible improved survival of patients with stage IV AJCC melanoma receiving SRL 172 immunotherapy: correlation with induction of increased levels of intracellular interleukin-2 in peripheral blood lymphocytes Ann Oncol 1999 10: 817–824

    Article  CAS  PubMed  Google Scholar 

  8. Heriot AG et al. Reduction in cytokine production in colorectal cancer patients: association with stage and reversal by resection Br J Cancer 2000 82: 1009–1012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Blades RA et al. Loss of HLA class I expression in prostate cancer: implications for immunotherapy Urology 1995 46: 681–687

    Article  CAS  PubMed  Google Scholar 

  10. Freeman SM et al. The ‘bystander effect’: tumor regression when a fraction of the tumor mass is genetically modified Cancer Res 1993 53: 5274–5283

    CAS  PubMed  Google Scholar 

  11. Vile RG et al. Systemic gene therapy of murine melanoma using tissue specific expression of the HSVtk gene involves an immune component Cancer Res 1994 54: 6228–6234

    CAS  PubMed  Google Scholar 

  12. Ramesh R et al. In vivo analysis of the ‘bystander effect’: a cytokine cascade Exp Hematol 1996 24: 829–838

    CAS  PubMed  Google Scholar 

  13. Alvarez RD, Curiel DT . A phase I study of recombinant adenovirus vector-mediated intraperitoneal delivery of herpes simplex virus thymidine kinase (HSV-TK) gene and intravenous ganciclovir for previously treated ovarian and extraovarian cancer patients Hum Gene Ther 1997 8: 597–613

    Article  CAS  PubMed  Google Scholar 

  14. Sterman DH et al. Adenovirus-mediated herpes simplex virus thymidine kinase/ganciclovir gene therapy in patients with localized malignancy: results of a phase I clinical trial in malignant mesothelioma Hum Gene Ther 1998 9: 1083–1092

    Article  CAS  PubMed  Google Scholar 

  15. Herman JR et al. In situ gene therapy for adenocarcinoma of the prostate: a phase I clinical trial Hum Gene Ther 1999 10: 1239–1249

    Article  CAS  PubMed  Google Scholar 

  16. Pettaway CA et al. Selection of highly metastatic variants of different human prostatic carcinomas using orthotopic implantation in nude mice Clin Cancer Res 1996 2: 1627–1636

    CAS  PubMed  Google Scholar 

  17. Stubbs M, Griffiths JR . Monitoring cancer by magnetic resonance Br J Cancer 1999 80 (Suppl. 1): 86–94

    PubMed  Google Scholar 

  18. Griffiths JR et al. 31P-NMR investigation of solid tumours in the living rat Biosci Reps 1981 1: 319–325

    Article  CAS  Google Scholar 

  19. Griffiths JR et al. 31P-NMR studies of a human tumour in situ Lancet 1983 i: 1435–1436

    Article  Google Scholar 

  20. deCertaines JD et al. In vivo 31P MRS of experimental tumours NMR Biomed 1993 6: 345–365

    Article  CAS  Google Scholar 

  21. Ross BD, Kim B, Davidson B . Assessment of ganciclovir toxicity to experimental intracranial gliomas following recombinant adenoviral mediated transfer of the herpes simplex virus thymidine kinase gene by magnetic resonance imaging and proton magnetic resonance spectroscopy Clin Cancer Res 1995 1: 651–657.

    CAS  PubMed  Google Scholar 

  22. Izquierdo M et al. Long term rat survival after malignant brain-tumor regression by retroviral gene-therapy Gene Therapy 1995 2: 66–69

    CAS  PubMed  Google Scholar 

  23. Namba H et al. Evaluation of the bystander effect in experimental brain tumors bearing herpes simplex virus-thymidine kinase gene by serial magnetic resonance imaging Hum Gene Ther 1996 7: 1847–1852

    Article  CAS  PubMed  Google Scholar 

  24. Stegman LD, Ben-Yoseph O, Freyer JP, Ross BD . In vivo 31P MRS evaluation of ganciclovir toxicity in C6 gliomas stably expressing the herpes simplex thymidine kinase gene NMR Biomed 1996 9: 364–368

    Article  CAS  PubMed  Google Scholar 

  25. Hakumäki JM et al. 1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: implications for the in vivo detection of apoptosis Nat. Med 1999 5: 1323–1327

    Article  PubMed  Google Scholar 

  26. Melcher A et al. Tumor immunogenicity is determined by the mechanism of cell death via induction of heat shock protein expression Nat. Med 1998 4: 581–587

    Article  CAS  PubMed  Google Scholar 

  27. Hrouda D et al. Mycobacterium vaccae (SRL172): a potential immunological adjuvant evaluated in rat prostate cancer Br J Urol 1998 82: 870–876

    Article  CAS  PubMed  Google Scholar 

  28. Hrouda D et al. Allogeneic whole tumour cell vaccination in the rat model of prostate cancer Br J Urd 2000 86: 742–748

    Article  CAS  Google Scholar 

  29. Vieweg J et al. Immunotherapy of prostate cancer in the Dunning rat model: use of cytokine gene modified tumor vaccines Cancer Res 1994 54: 1760–1765

    CAS  PubMed  Google Scholar 

  30. Sanda MG et al. Demonstration of a rational strategy for human prostate cancer gene therapy J Urol 1994 151: 622–628

    Article  CAS  PubMed  Google Scholar 

  31. Eastham JA et al. Prostate cancer gene therapy: herpes simplex virus thymidine kinase gene transduction followed by ganciclovir in mouse and human prostate cancer models Hum Gene Ther 1996 7: 515–523

    Article  CAS  PubMed  Google Scholar 

  32. Vile RG et al. Generation of an anti-tumour immune response in a non-immunogenic tumour: HSVtk killing in vivo stimulates a mononuclear cell infiltrate and a Th1-like profile of intratumoural cytokine expression Int J Cancer 1997 71: 267–274

    Article  CAS  PubMed  Google Scholar 

  33. Vile RG, Diaz RM, Castleden S, Chong H . Targeted gene therapy for cancer: herpes simplex virus thymidine kinase gene-mediated cell killing leads to anti-tumour immunity that can be augmented by co-expression of cytokines in the tumour cells Biochem Soc Trans 1997 25: 717–722

    Article  CAS  PubMed  Google Scholar 

  34. Steen RG et al. 31P NMR spectroscopic and near infrared spectrophotometric studies of effects of anesthetics on in vivo RIF-1 tumors. Relationship to tumor radiosensitivity NMR Biomed 1989 2: 87–92

    Article  CAS  PubMed  Google Scholar 

  35. Rodriguez LM et al. 31P nuclear magnetic resonance spectroscopy studies of the response of rat mammary tumours to endocrine therapy Cancer Res 1988 48: 89–93

    Google Scholar 

  36. Negendank WG . Studies on human tumours by MRS: a review NMR Biomed 1992 5: 303–324

    Article  CAS  PubMed  Google Scholar 

  37. Ruiz-Cabello J, Cohen JS . Phospholipid metabolites as indicators of cancer cell function NMR Biomed 1992 5: 226–233

    Article  CAS  PubMed  Google Scholar 

  38. Ordidge RJ, Connelly A, Lohman JAB . Image-selected in vivo spectroscopy (ISIS). A new technique for spatially selective NMR spectroscopy J Magn Reson 1986 66: 283–294

    CAS  Google Scholar 

  39. van der Veen JWC, de Beer R, Luyten PR, Van Ormondt D . Accurate quantification of in vivo P-31 NMR signals using the variable projection method and prior knowledge Magn Res Med 1988 6: 92–98

    Article  CAS  Google Scholar 

Download references

Acknowledgements

JD Eaton was supported by the Swire Group. MJA Perry was supported by the British Urological Foundation and Onyvax Ltd. RA Mazucco was supported by a grant from the Cancer Research Campaign UK (CRC) SP1971/0404. SM Todryk was supported by Onyvax Ltd. We are grateful to Dr R Vile (Mayo Clinic, Rochester, USA) for providing the HSVtk retroviral packaging line, and to Dr H Pandha and Dr M Stubbs (St George's Hospital Medical School) for very helpful discussion.

Author information

Author notes

  1. J D Eaton and M J A Perry: The first two authors contributed equally to this study

Authors and Affiliations

  1. Division of Oncology, St George's Hospital Medical School, London, UK

    J D Eaton, M J A Perry, S M Todryk & A G Dalgleish

  2. Department of Urology, St George's Hospital, London, UK

    J D Eaton, M J A Perry & R S Kirby

  3. Department of Biochemistry and Immunology, St George's Hospital Medical School, London, UK

    R A Mazucco & J R Griffiths

Authors
  1. J D Eaton
    View author publications

    Search author on:PubMed Google Scholar

  2. M J A Perry
    View author publications

    Search author on:PubMed Google Scholar

  3. S M Todryk
    View author publications

    Search author on:PubMed Google Scholar

  4. R A Mazucco
    View author publications

    Search author on:PubMed Google Scholar

  5. R S Kirby
    View author publications

    Search author on:PubMed Google Scholar

  6. J R Griffiths
    View author publications

    Search author on:PubMed Google Scholar

  7. A G Dalgleish
    View author publications

    Search author on:PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eaton, J., Perry, M., Todryk, S. et al. Genetic prodrug activation therapy (GPAT) in two rat prostate models generates an immune bystander effect and can be monitored by magnetic resonance techniques. Gene Ther 8, 557–567 (2001). https://doi.org/10.1038/sj.gt.3301432

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/sj.gt.3301432

Keywords

This article is cited by

Search

Advanced search

Quick links