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Differential transduction efficacy of adenoviruses containing NGR within cyclic and linear sequences in HI loop of fiber protein to aminopeptidase N and alpha V beta 3 integrins

The aim of this work was to design an gene therapy applicable Ad vector retargeted on the APN molecule expressed on angiogenetic cells via insertion of specific NGR motiv (asparagine-glycine-arginine) into the HI loop of a fiber protein. On the rhabdomyosarcoma (RD) cell line, which expresses APN but only very low levels of α vβ 3 integrin and CAR receptor, all NGR-bearing Ad we have constructed exhibited moderately increased attachment and transduction efficacy in comparison to wild type virus. NGR-bearing Ad containing cyclic motifs were more efficient than those containing linear ones. The increased transduction efficacy of NGR-bearing Ads was completely abolished by the APN-specific peptide CNGRC and the integrin-specific peptide CRGDC. By measuring transduction efficacy on human laryngeal carcinoma cells with graded expression of α vβ 3 integrin, we found that NGR-bearing Ad bound weakly to α vβ 3 integrin. Treatment of RD cells with TGF-1 up-regu lated APN and increased transduction efficacy with Ad bearing NGR within cyclic sequence while this effect is less pronounced for Ad bearing NGR within linear sequence and is likely to be predominantly consequence of moderately increased expression of α vβ 3 integrin. Both, APN and α vβ 3 integrin are up regulated in endothelial cells in angiogenesis, therefore NGR-bearing adenoviruses could be suitable vectors for tumor gene therapy.

adenovirus; gene retargeting; aminopeptidase n

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