Nature Medicine
5, 1052 - 1056 (1999)
doi:10.1038/12491
Genetic capsid modifications allow efficient re-targeting of adeno-associated
virus type 2Anne Girod1, Martin Ried1, Christiane Wobus2, Harald Lahm1, Kristin Leike1, Jürgen Kleinschmidt2, Gilbert Deléage3
& Michael Hallek1, 41
Laboratorium für Molekulare Biologie, Genzentrum Medizinische Klinik III, Klinikum Gro hadern, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strae 25, D-81377 München, Germany
4
Medizinische Klinik III, Klinikum Grohadern, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strae 25, D-81377 München, Germany
2
Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany
3
Pôle BioInformatique Lyonnais, Laboratoire de Conformation des Protéines, Institut de Biologie et Chimie des Protéines, UPR412-CNRS, F-69367 Lyon Cedex 07, France
Correspondence should be addressed to Michael Hallek hallek@lmb.uni-muenchen.deThe 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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