Naslov
Disulfide bond formation in NGR fiber-modified adenovirus is essential for retargeting to aminopeptidase N

Autori
Majhen, Dragomira ; Gabrilovac, Jelka ; Eloit, Marc ; Richardson, Jeniffer ; Ambriović-Ristov, Andreja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
The 8th International Adenovirus Meeting: Program and Abstracts / - Zürich, 2006

Skup
8th International Adenovirus Meeting

Mjesto i datum
Zürich, Švicarska, 30.08.2006. - 02.09.2006

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
adenovirus; gene retargeting; aminopeptidase N

Sažetak
: Peptides that permit targeting to specific cell types may be identified by phage display. The peptide motif NGR (asparagine-glycine-arginine) is known to bind to aminopeptidase N (APN) expressed on the endothelial cells of angiogenic vasculature, representing a target molecule for tumor gene therapy aimed at inhibition of angiogenesis. The goal of this work was to design an Adenovirus vector type 5 (Ad5) retargeted on the APN molecule via incorporation of specific NGR-containing ligands into the HI loop of a fiber protein and to examine the effect of linear and cyclic sequences on its targeting properties. We constructed four replication defective adenoviruses bearing cyclic or linear NGR-containing sequences and showed that these insertions did not affect structure or incorporation of the fiber protein in viral particles. We have shown on the rhabdomyosarcoma (RD) cell line which expresses APN, and a model composed of a human laryngeal carcinoma cell line (HEp2) derived stably transfected cell lines with graded expression levels of the alpha v beta 3, that cellular receptor(s) for adenoviruses bearing cyclic or linear NGR-containing sequences were not identical. Ads containing NGR within potentially cyclic sequences flanked by cysteines retargeted viruses mainly to APN, while Ads containing NGR within linear sequences not containing cysteines retargeted Ads mainly to alpha v beta 3 integrin, albeit with a lower affinity. Finally, we show evidence that disulfide bond formation within an Ad bearing the CDCNGRCFC sequence is essential for retargeting to APN, suggesting that this sequence does indeed assume a cyclic structure which facilitates NGR binding to APN. Therefore, our study underscores the importance of cysteine residues flanking targeting peptides for not only affinity but also specificity of the retargeted Ad.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA