Adeno-associated virus capsids displaying immunoglobulin-binding domains permit antibody-mediated vector retargeting to specific cell surface receptors.
Recombinant adeno-associated virus type 2 (rAAV2) is a promising vector for human somatic gene therapy. However, its broad host range is a disadvantage for some applications, because it reduces the specificity of the gene transfer. To overcome this limitation, we sought to create a versatile rAAV vector targeting system which would allow us to redirect rAAV binding to specific cell surface receptors by simple coupling of different ligands to its capsid. For this purpose, an immunoglobulin G (IgG) binding domain of protein A, Z34C, was inserted into the AAV2 capsid at amino acid position 587. The resulting AAV2-Z34C mutants could be packaged and purified to high titers and bound to IgG molecules. rAAV2-Z34C vectors coupled to antibodies against CD29 (beta(1)-integrin), CD117 (c-kit receptor), and CXCR4 specifically transduced distinct human hematopoietic cell lines. In marked contrast, no transduction was seen in the absence of antibodies or in the presence of specific blocking reagents. These results demonstrate for the first time that an immunoglobulin binding domain can be inserted into the AAV2 capsid and coupled to various antibodies, which mediate the retargeting of rAAV vectors to specific cell surface receptors.
Ried
Martin U
MU
Laboratorium für Molekulare Biologie, Genzentrum, Germany.
Girod
Anne
A
Leike
Kristin
K
Büning
Hildegard
H
Hallek
Michael
M
D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism*; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D006412:Hematopoietic Stem Cells; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism*; D019012:Integrin beta1 / Q000378:metabolism; D008024:Ligands; D016254:Mutagenesis, Insertional; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011817:Rabbits; D019718:Receptors, CXCR4 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014161:Transduction, Genetic
D000914:Antibodies, Viral; D007136:Immunoglobulins; D019012:Integrin beta1; D008024:Ligands; D019718:Receptors, CXCR4; D011956:Receptors, Cell Surface; D019009:Proto-Oncogene Proteins c-kit