title: Genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2.
abstract: The human parvovirus adeno-associated virus type 2 (AAV2) has many features that make it attractive as a vector for gene therapy. 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-receptors. 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.
authors.affiliation: Laboratorium für Molekulare Biologie, Genzentrum Medizinische Klinik III, Klinikum Grobetahadern, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strabetae 25, D-81377 München, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Girod
authors.forename: M
authors.initials: M
authors.lastname: Ried
authors.forename: C
authors.initials: C
authors.lastname: Wobus
authors.forename: H
authors.initials: H
authors.lastname: Lahm
authors.forename: K
authors.initials: K
authors.lastname: Leike
authors.forename: J
authors.initials: J
authors.lastname: Kleinschmidt
authors.forename: G
authors.initials: G
authors.lastname: Deléage
authors.forename: M
authors.initials: M
authors.lastname: Hallek
mesh_terms: D000595:Amino Acid Sequence; D001667:Binding, Competitive; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D016023:Integrins / Q000378:metabolism; D007797:Laminin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D011991:Receptors, Virus / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic / Q000187:drug effects / Q000235:genetics; D014407:Tumor Cells, Cultured; D019065:Virus Assembly
chemical_list: D016023:Integrins; D007797:Laminin; D008024:Ligands; D009842:Oligopeptides; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C047981:arginyl-glycyl-aspartic acid; D006493:Heparin