Pronounced antitumor efficacy by extracellular activation of a doxorubicin-glucuronide prodrug after adenoviral vector-mediated expression of a human antibody-enzyme fusion protein
| Autor: | Michelle de Graaf, Epie Boven, Ida H van der Meulen-Muileman, D. Oosterhoff, Hidde J. Haisma, Winald R. Gerritsen, Herbert M. Pinedo |
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| Rok vydání: | 2004 |
| Předmět: |
Genetic enhancement
Recombinant Fusion Proteins Genetic Vectors Antineoplastic Agents Glucuronates Biology medicine.disease_cause Antibodies Viral vector Adenoviridae chemistry.chemical_compound Mice Antigens Neoplasm Cell Line Tumor Neoplasms Spheroids Cellular Genetics medicine Animals Humans Doxorubicin Prodrugs Molecular Biology Glucuronidase Ovarian Neoplasms Gene Transfer Techniques Prodrug Epithelial Cell Adhesion Molecule Flow Cytometry Fusion protein Molecular biology Xenograft Model Antitumor Assays chemistry Cell culture Molecular Medicine Female Growth inhibition Cell Adhesion Molecules medicine.drug Protein Binding |
| Zdroj: | Human gene therapy. 15(3) |
| ISSN: | 1043-0342 |
| Popis: | Tumor-specific activation of the glucuronide prodrug of doxorubicin, N-[4-doxorubicin-N-carbonyl( oxymethyl) phenyl]-O-beta-glucuronyl carbamate (DOX-GA3), by beta-glucuronidase present in necrotic tumor areas might be improved after transduction of tumor cells to secrete a targeted form of beta-glucuronidase. To that end, we constructed an adenovirus vector, designated Ad/C28-GUSh, encoding human beta-glucuronidase fused to a human single-chain Fv ( scFv) against the epithelial cell adhesion molecule ( EpCAM), C28, and preceded by a signal sequence for secretion. Antibody specificity and enzyme activity were retained in the fusion protein secreted by tumor cells infected with Ad/C28-GUSh. Diffusion of fusion protein from transduced tumor cells within MCF-7 multicellular spheroids was visualized by immunohistochemistry. Treatment of spheroids with Ad/C28-GUSh and DOX-GA3 resulted in growth inhibition comparable to treatment with doxorubicin alone. Treatment of well-established FMa human ovarian cancer xenografts with intravenous injection of DOX-GA3 ( 500 mg/kg) resulted in a tumor volume-doubling time of 23.8 days compared to 8.0 days for phosphate-buffered saline (PBS)-treated mice. Intratumoral administration of Ad/C28-GUSh before DOX-GA3 enhanced the growth inhibition and increased the tumor volume-doubling time to 43.1 days (p |
| Databáze: | OpenAIRE |
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