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Tumor cell targeted gene delivery by adenovirus 5 vectors carrying knobless fibers with antibody-binding domains

Gene Therapy volume 12, pages 211–224 (2005)Cite this article

Abstract

Most human carcinoma cell lines lack the high-affinity receptors for adenovirus serotype 5 (Ad5) at their surface and are nonpermissive to Ad5. We therefore tested the efficiency of retargeting Ad5 to alternative cellular receptors via immunoglobulin (Ig)-binding domains inserted at the extremity of short-shafted, knobless fibers. The two recombinant Ad5's constructed, Ad5/R7-Zwt-Zwt and Ad5/R7-C2-C2, carried tandem Ig-binding domains from Staphylococcal protein A (abbreviated Zwt) and from Streptococcal protein G (C2), respectively. Both viruses bound their specific Ig isotypes with the expected affinity. They transduced human carcinoma cells independently of the CAR pathway, via cell surface receptors targeted by specific monoclonal antibodies, that is, EGF-R on A549, HT29 and SW1116, HER-2/neu on SK-OV-3 and SK-BR-3, CA242 (epitope recognized by the monoclonal antibody C242) antigen on HT29 and SW1116, and PSMA (prostate-specific membrane antigen) expressed on HEK-293 cells, respectively. However, Colo201 and Colo205 cells were neither transduced by targeting CA242 or EGF-R nor were LNCaP cells transduced by targeting PSMA. Our results suggested that one given surface receptor could mediate transduction of certain cells but not others, indicating that factors and steps other than cell surface expression and virus–receptor interaction are additional determinants of Ad5-mediated transduction of tumor cells. Using penton base RGD mutants, we found that one of these limiting steps was virus endocytosis.

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Acknowledgements

We are grateful to Simone Peyrol and Christel Mathias (Centre Commun d’Imagerie, Faculté de Médecine Laennec) for their help in electron microscopy. We acknowledge the Swegene Centre of Cellular Imaging at Gothenburg University for the use of imaging equipment. This work was financially supported by grants from Got-a-Gene AB, Stena Center, Gothenburg, Sweden, from the Association Françai'se contre les Myopathies (AFM), Institut de Myologie, Paris (Subvention AFM/Groupe E/2003/11845-9481), and from the Association Vaincre la Mucoviscidose (VLM), Paris, France.

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  1. P Henning and K M E Andersson: These authors contributed equally to this work

Authors and Affiliations

  1. Got-a-Gene AB, Gothenburg, Sweden

    P Henning, K M E Andersson, K Frykholm, A Ali, M K Magnusson & L Lindholm

  2. Department of Medical Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden

    P Henning & L Lindholm

  3. Department of Biotechnology, Royal Institute of Technology, Alba Nova University Center, Stockholm, Sweden

    P-Å Nygren

  4. Laboratoire de Virologie et Pathogenèse Virale, Faculté de Médecine RTH Laennec, Lyon, France

    O Granio, S S Hong & P Boulanger

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Henning, P., Andersson, K., Frykholm, K. et al. Tumor cell targeted gene delivery by adenovirus 5 vectors carrying knobless fibers with antibody-binding domains. Gene Ther 12, 211–224 (2005). https://doi.org/10.1038/sj.gt.3302408

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