TY - JOUR
T1 - Targeting of Adenovirus via Genetic Modification of the Viral Capsid Combined with a Protein Bridge
AU - Korokhov, Nikolay
AU - Mikheeva, Galina
AU - Krendelshchikov, Alexander
AU - Belousova, Natalya
AU - Simonenko, Vera
AU - Krendelshchikova, Valentina
AU - Pereboev, Alexander
AU - Kotov, Alexander
AU - Kotova, Olga
AU - Triozzi, Pierre L.
AU - Aldrich, Wayne A.
AU - Douglas, Joanne T.
AU - Lo, Kin Ming
AU - Banerjee, Papia T.
AU - Gillies, Stephen D.
AU - Curiel, David T.
AU - Krasnykh, Victor
PY - 2003/12
Y1 - 2003/12
N2 - A potential barrier to the development of genetically targeted adenovirus (Ad) vectors for cell-specific delivery of gene therapeutics lies in the fact that several types of targeting protein ligands require posttranslational modifications, such as the formation of disulfide bonds, which are not available to Ad capsid proteins due to their nuclear localization during assembly of the virion. To overcome this problem, we developed a new targeting strategy, which combines genetic modifications of the Ad capsid with a protein bridge approach, resulting in a vector-ligand targeting complex. The components of the complex associate by virtue of genetic modifications to both the Ad capsid and the targeting ligand. One component of this mechanism of association, the Fc-binding domain of Staphylococcus aureus protein A, is genetically incorporated into the Ad fiber protein. The ligand is comprised of a targeting component fused with the Fc domain of immunoglobulin, which serves as a docking moiety to bind to these genetically modified fibers during the formation of the Ad-ligand complex. The modular design of the ligand solves the problem of structural and biosynthetic compatibility with the Ad and thus facilitates targeting of the vector to a variety of cellular receptors. Our study shows that targeting ligands incorporating the Fc domain and either an anti-CD40 single-chain antibody or CD40L form stable complexes with protein A-modified Ad vectors, resulting in significant augmentation of gene delivery to CD40-positive target cells. Since this gene transfer is independent of the expression of the native Ad5 receptor by the target cells, this strategy results in the derivation of truly targeted Ad vectors suitable for tissue-specific gene therapy.
AB - A potential barrier to the development of genetically targeted adenovirus (Ad) vectors for cell-specific delivery of gene therapeutics lies in the fact that several types of targeting protein ligands require posttranslational modifications, such as the formation of disulfide bonds, which are not available to Ad capsid proteins due to their nuclear localization during assembly of the virion. To overcome this problem, we developed a new targeting strategy, which combines genetic modifications of the Ad capsid with a protein bridge approach, resulting in a vector-ligand targeting complex. The components of the complex associate by virtue of genetic modifications to both the Ad capsid and the targeting ligand. One component of this mechanism of association, the Fc-binding domain of Staphylococcus aureus protein A, is genetically incorporated into the Ad fiber protein. The ligand is comprised of a targeting component fused with the Fc domain of immunoglobulin, which serves as a docking moiety to bind to these genetically modified fibers during the formation of the Ad-ligand complex. The modular design of the ligand solves the problem of structural and biosynthetic compatibility with the Ad and thus facilitates targeting of the vector to a variety of cellular receptors. Our study shows that targeting ligands incorporating the Fc domain and either an anti-CD40 single-chain antibody or CD40L form stable complexes with protein A-modified Ad vectors, resulting in significant augmentation of gene delivery to CD40-positive target cells. Since this gene transfer is independent of the expression of the native Ad5 receptor by the target cells, this strategy results in the derivation of truly targeted Ad vectors suitable for tissue-specific gene therapy.
UR - http://www.scopus.com/inward/record.url?scp=10744228126&partnerID=8YFLogxK
U2 - 10.1128/JVI.77.24.12931-12940.2003
DO - 10.1128/JVI.77.24.12931-12940.2003
M3 - Article
C2 - 14645549
AN - SCOPUS:10744228126
SN - 0022-538X
VL - 77
SP - 12931
EP - 12940
JO - Journal of virology
JF - Journal of virology
IS - 24
ER -