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A cathepsin-cleavage site between the adenovirus capsid protein IX and a tumor-targeting ligand improves targeted transduction

Gene Therapy volume 19, pages 899–906 (2012)Cite this article

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Abstract

Human adenoviruses have a great potential as anticancer agents. One strategy to improve their tumor-cell specificity and anti-tumor efficacy is to include tumor-specific targeting ligands in the viral capsid. This can be achieved by fusion of polypeptide-targeting ligands with the minor capsid protein IX. Previous research suggested that protein IX-mediated targeting is limited by inefficient release of protein IX-fused ligands from their cognate receptors in the endosome. This thwarts endosomal escape of the virus particles. Here we describe that the targeted transduction of tumor cells is augmented by a cathepsin-cleavage site between the protein IX anchor and the HER2/neu-binding ZH Affibody molecule as ligand. The cathepsin-cleavage site did not interfere with virus production and incorporation of the Affibody molecules in the virus capsid. Virus particles harboring the cleavable protein IX–ligand fusion in their capsid transduced the HER2/neu-positive SKOV-3 ovarian carcinoma cells with increased efficiency in monolayer cultures, three-dimensional spheroid cultures and in SKOV-3 tumors grown on the chorioallantoic membrane of embryonated chicken eggs. These data show that inclusion of a cathepsin-cleavage sequence between protein IX and a high-affinity targeting ligand enhances targeted transduction. This modification further augments the applicability of protein IX as an anchor for coupling tumor-targeting ligands.

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Acknowledgements

We thank Masato Yamamoto (Department of Surgery, University of Minnesota, MN, USA) for providing us with the plasmid pShuttle-ΔE3-ADP-EGFP-F2. In addition, Ivo Que (Department of Endocrinology, Leiden University Medical Center, Leiden, Netherlands) is gratefully acknowledged for precious technical assistance in the biofluorescence experiments. This work was supported by the European Union through the 6th Framework Program GIANT (Contract No. 512087).

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

    J de Vrij, I J C Dautzenberg, S K van den Hengel, T G Uil, S J Cramer, J Vellinga, C S Verissimo, D Koppers-Lalic & R C Hoeben

  2. Got-A-Gene AB, Kullavik, Sweden

    M K Magnusson & L Lindholm

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Correspondence to R C Hoeben.

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de Vrij, J., Dautzenberg, I., van den Hengel, S. et al. A cathepsin-cleavage site between the adenovirus capsid protein IX and a tumor-targeting ligand improves targeted transduction. Gene Ther 19, 899–906 (2012). https://doi.org/10.1038/gt.2011.162

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