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Genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2

Nature Medicine volume 5, pages 1052–1056 (1999)Cite this article

An Erratum to this article was published on 01 December 1999

Abstract

The human parvovirus adeno-associated virus type 2 (AAV2) has many features that make it attractive as a vector for gene therapy1,2. However, the broad host range of AAV2 might represent a limitation for some applications in vivo, because recombinant AAV vector (rAAV)-mediated gene transfer would not be specific for the tissue of interest. This host range is determined by the binding of the AAV2 capsid to specific cellular receptors and/or co-receptors3,4,5,6. The tropism of AAV2 might be changed by genetically introducing a ligand peptide into the viral capsid, thereby redirecting the binding of AAV2 to other cellular receptors. We generated six AAV2 capsid mutants by inserting a 14-amino-acid targeting peptide, L14, into six different putative loops of the AAV2 capsid protein identified by comparison with the known three-dimensional structure of canine parvovirus. All mutants were efficiently packaged. Three mutants expressed L14 on the capsid surface, and one efficiently infected wild-type AAV2-resistant cell lines that expressed the integrin receptor recognized by L14. The results demonstrate that the AAV2 capsid tolerates the insertion of a nonviral ligand sequence. This might open new perspectives for the design of targeted AAV2 vectors for human somatic gene therapy.

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Figure 1: Delineation of insertion sites for a ligand peptide in the AAV2 capsid protein.
Figure 2: Presentation and functionality of the L14 sequence.
Figure 3: Transduction of B16F10 cells.

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Acknowledgements

We thank I. Renner-Müller and E. Wolf for their help and providing their animal facilities to produce rabbit antiserum against L14. This work was supported by grants from the Wilhelm-Sanderstiftung, Bayerische Forschungsstiftung, and the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 455) (M.H. and A.G.) and stipends of the Studienstiftung des Deutschen Volkes and the Fond der Chemischen Industrie (BMBF) (M.R.).

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Authors and Affiliations

  1. Laboratorium für Molekulare Biologie, Genzentrum Medizinische Klinik III, Klinikum Groβhadern, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straβe 25, München, D-81377, Germany

    Anne Girod, Martin Ried, Harald Lahm, Kristin Leike & Michael Hallek

  2. Medizinische Klinik III, Klinikum Groβhadern, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straβe 25, München, D-81377, Germany

    Michael Hallek

  3. Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Im Neuenheimer Feld 242, Heidelberg, D-69120, Germany

    Christiane Wobus & Jürgen Kleinschmidt

  4. Pôle BioInformatique Lyonnais, Laboratoire de Conformation des Protéines, Institut de Biologie et Chimie des Protéines, UPR412-CNRS 07, Lyon, F-69367 Cedex, France

    Gilbert Deléage

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  1. Anne Girod
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Correspondence to Michael Hallek.

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Girod, A., Ried, M., Wobus, C. et al. Genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2. Nat Med 5, 1052–1056 (1999). https://doi.org/10.1038/12491

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