Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy.
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 beta-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 beta-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.
Heine
D
D
Institute of Molecular Biology and Tumor Research (IMT), Philipps-University Marburg, Emil-Mannkopff-Strasse 2, D-35033 Marburg, Germany.
Müller
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R
Brüsselbach
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S
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002822:Choriocarcinoma / Q000473:pathology / Q000628:therapy; D004317:Doxorubicin / Q000031:analogs & derivatives / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods*; D005965:Glucuronates / Q000378:metabolism*; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology / Q000628:therapy; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D011355:Prodrugs / Q000378:metabolism*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D000903:Antibiotics, Antineoplastic; D005965:Glucuronates; C114198:N-(4-glucuronyl-3-nitrobenzyloxycarbonyl)doxorubicin; D011355:Prodrugs; D004317:Doxorubicin; D005966:Glucuronidase