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.

  • Acquired Diseases
  • Published:

Secreted human β-glucuronidase: a novel tool for gene-directed enzyme prodrug therapy

Gene Therapy volume 7, pages 224–231 (2000)Cite this article

Abstract

A major problem of tumor gene therapy is the low transduction efficiency of the currently available vectors. One way to circumvent this problem is the delivery of therapeutic genes encoding intracellular enzymes for the conversion of a prodrug to a cytotoxic drug which can then spread to neighboring non-transduced cells (bystander effect). One possibility to improve the bystander effect could be the extracellular conversion of a hydrophilic prodrug to a lipophilic, cell-permeable cytotoxic drug. Toward this end, we have used a secreted form of the normally lysosomal human β-glucuronidase (s-βGluc) to establish an extracellular cytotoxic effector system that converts an inactivated glucuronidated derivative of doxorubicin (HMR 1826) to the cytotoxic drug. We demonstrate that s-βGluc-transduced tumor cells convert HMR 1826 to doxorubicin which is taken up by both transduced and non-transduced cells. s-βGluc in combination with HMR 1826 efficiently induces tumor cell killing both in cell culture and in vivo. This effect is mediated through a pronounced bystander effect of the generated cytotoxic drug. Most notably, this gene therapeutic strategy is shown to be clearly superior to conventional chemotherapy with doxorubicin.

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

Access options

Buy this article

  • Purchase on Springer Link
  • 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

Similar content being viewed by others

References

  1. Ford JM, Hait WN . Pharmacology of drugs that alter multidrug resistance in cancer Pharmacol Rev 1990 42: 155–199

    CAS  PubMed  Google Scholar 

  2. Culver KW et al. In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors (see comments) Science 1992 256: 1550–1552

    Article  CAS  PubMed  Google Scholar 

  3. Mullen CA, Kilstrup M, Blaese RM . Transfer of the bacterial gene for cytosine deaminase to mammalian cells confers lethal sensitivity to 5-fluorocytosine; a negative selections sytem Proc Natl Acad Sci USA 1992 89: 33–37

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Huber BE et al. In vivo antitumor activity of 5-fluorocytosine on human colorectal carcinoma cells genetically modified to express cytosine deaminase Cancer Res 1993 53: 4619–4626

    CAS  PubMed  Google Scholar 

  5. Bridgewater JA et al. Expression of the bacterial nitroreductase enzyme in mammalian cells renders them selectively sensitive to killing by the prodrug CB1954 (published erratum appears in Eur J Cancer 1996; 32A: 186) Eur J Cancer 1995 31A: 2362–2370

    Article  CAS  PubMed  Google Scholar 

  6. Marais R et al. A cell surface tethered enzyme improves efficiency in gene-directed enzyme prodrug therapy Nature Biotech 1997 15: 1373–1377

    Article  CAS  Google Scholar 

  7. Paigen K . Mammalian beta-glucuronidase: genetics, molecular biology, and cell biology Prog Nucleic Acid Res Mol Biol 1989 37: 155–205

    Article  CAS  PubMed  Google Scholar 

  8. Rosenfeld MG et al. Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution J Cell Biol 1982 93: 135–143

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Jain S et al. Structure of human beta-glucuronidase reveals candidate lysosomal targeting and active-site motifs Nat Struct Biol 1996 3: 375–381

    Article  CAS  PubMed  Google Scholar 

  10. Bosslet K, Czech J, Hoffmann D . A novel one-step tumor-selective prodrug activation system Tumor Targetting 1995 1: 45–50

    CAS  Google Scholar 

  11. Barabas K, Sizensky JA, Faulk WP . Transferrin conjugates of adriamycin are cytotoxic without intercalating nuclear DNA J Biol Chem 1992 267: 9437–9442

    CAS  PubMed  Google Scholar 

  12. Fick J et al. The extent of heterocellular communication mediated by gap junctions is predictive of bystander tumor cytotoxicity in vitro Proc Natl Acad Sci USA 1995 92: 11071–11075

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mesnil M et al. Bystander killing of cancer cells by herpes simplex virus thymidine kinase gene is mediated by connexins Proc Natl Acad Sci USA 1996 93: 1831–1835

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Freeman SM et al. The ‘bystander effect’: tumor regression when a fraction of the tumor mass is genetically modified Cancer Res 1993 53: 5274–52783

    CAS  PubMed  Google Scholar 

  15. Bridgewater JA et al. The bystander effect of the nitro-reductase/CB1954 enzyme/prodrug system is due to a cell-permeable metabolite Hum Gene Ther 1997 8: 709–717

    Article  CAS  PubMed  Google Scholar 

  16. Vaupel P, Kallinowski F, Okunieff P . Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review Cancer Res 1989 49: 6449–6465

    CAS  PubMed  Google Scholar 

  17. Tannock IF, Rotin D . Acid pH in tumors and its potential for therapeutic exploitation Cancer Res 1989 49: 4373–4384

    CAS  PubMed  Google Scholar 

  18. Lougerstay-Madec R et al. Synthesis of self-immolative glucuronide-based prodrugs of a phenol mustard Anticancer Drug Des 1998 13: 995–1007

    CAS  PubMed  Google Scholar 

  19. Bakina E, Wu Z, Rosenblum M, Farquhar D . Intensely cytotoxic anthracycline prodrugs: glucuronides J Med Chem 1997 40: 4013–4018

    Article  CAS  PubMed  Google Scholar 

  20. Abina MA et al. Lac Z gene transfer into tumor cells abrogates tumorigenicity and protects mice against the development of further tumors Gene Therapy 1996 3: 212–216

    CAS  PubMed  Google Scholar 

  21. Van Ginkel FW et al. Intratracheal gene delivery with adenoviral vector induces elevated systemic IgG and mucosal IgA antibodies to adenovirus and beta-galactosidase Hum Gene Ther 1995 6: 895–903

    Article  CAS  PubMed  Google Scholar 

  22. 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 

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

    Article  CAS  PubMed  Google Scholar 

  24. Barba D, Hardin J, Sadelain M, Gage FH . Development of anti-tumor immunity following thymidine kinase-mediated killing of experimental brain tumors Proc Natl Acad Sci USA 1994 91: 4348–4352

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Mullen CA, Coale MM, Lowe R, Blaese RM . Tumors expressing the cytosine deaminase suicide gene can be eliminated in vivo with 5-glurocytosine and induce protective immunity to wild-type tumor Cancer Res 1994 54: 1503–1506

    CAS  PubMed  Google Scholar 

  26. Oshima A et al. Cloning, sequencing, and expression of cDNA for human beta-glucuronidase Proc Natl Acad Sci USA 1987 84: 685–689

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Riechmann L, Clark M, Waldmann H, Winter G . Reshaping human antibodies for therapy Nature 1988 332: 323–327

    Article  CAS  PubMed  Google Scholar 

  28. Graham FL et al. A new technique for the assay of infectivity of human adenovirus 5 DNA Virology 1973 52: 456–467

    Article  CAS  PubMed  Google Scholar 

  29. Bosslet K, Czech J, Hoffmann D . Tumor-selective prodrug activation by fusion protein-mediated catalysis Cancer Res 1994 54: 2151–2159

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Klaus Bosslet for useful discussions, to Claudia Cybon and Tina Stroh for excellent technical assistance, to A Wellstein for the JEG-3 cell line and to M Krause for synthesis of oligonucleotides.

Author information

Authors and Affiliations

  1. Institute of Molecular Biology and Tumor Research (IMT), Philipps-University Marburg, Germany

    D Weyel, R Müller & S Brüsselbach

  2. Hoechst Marion Roussel Deutschland, Marburg, Germany

    H-H Sedlacek

Authors
  1. D Weyel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H-H Sedlacek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Brüsselbach
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Weyel, D., Sedlacek, HH., Müller, R. et al. Secreted human β-glucuronidase: a novel tool for gene-directed enzyme prodrug therapy. Gene Ther 7, 224–231 (2000). https://doi.org/10.1038/sj.gt.3301072

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301072

Keywords

This article is cited by

Search

Advanced search

Quick links