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Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy

Gene Therapy volume 8, pages 1005–1010 (2001)Cite this article

Abstract

Prodrug conversion is a promising approach to cytotoxic gene therapy if an efficient transfer of the generated drug to adjacent cells can be achieved. To maximize the efficacy of this strategy we sought to develop a system that is based on a human enzyme, acts extracellularly yet in close vicinity of the transduced cell and can be used with multiple prodrugs. Results obtained with a secreted version of human β-glucuronidase suggested that this enzyme could be a suitable candidate, although a more stringent retention of the enzyme at the site of the producer cell, such as its attachment to the cell surface, would be desirable. Here, we show that the fusion of the transmembrane domain of the human PDGF receptor to a C-terminally truncated form of human β-glucuronidase results in its surface accumulation at high steady-state levels. Using a doxorubicin prodrug, we demonstrate that this GDEPT system produces a strong bystander effect and has potent antitumor activity in vivo.

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Acknowledgements

We are grateful to Prof HH Sedlacek for prodrug HMR1826 and monoclonal antibodies to β-glucuronidase, to Profs A Wellstein and K Havemann for JEG-3 and A549 cells, respectively, and to Imme Krüger, Tina Stroh, Julia Dick and Claudia Cybon for excellent technical assistance. This work is supported by a grant from the DFG to SB (BR 1857/2).

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  1. Institute of Molecular Biology and Tumor Research (IMT), Philipps-University Marburg, Emil-Mannkopff-Strasse 2, Marburg, D-35033, Germany

    D Heine, R Müller & S Brüsselbach

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  1. D Heine
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  2. R Müller
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  3. S Brüsselbach
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Heine, D., Müller, R. & Brüsselbach, S. Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy. Gene Ther 8, 1005–1010 (2001). https://doi.org/10.1038/sj.gt.3301474

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