Targeted adenovirus mediated inhibition of NF-κB-dependent inflammatory gene expression in endothelial cells in vitro and in vivo

• Published in: Journal of controlled release 2013-02, Vol.166 (1), p.57-65
• Main Authors: Kułdo, J.M., Ásgeirsdóttir, S.A., Zwiers, P.J., Bellu, A.R., Rots, M.G., Schalk, J.A.C., Ogawara, K.I., Trautwein, C., Banas, B., Haisma, H.J., Molema, G., Kamps, J.A.A.M.
• Format: Article
• Language: English
• Subjects: Adenoviral gene therapy; Adenoviridae; Adenoviridae - genetics; Adenovirus; adhesion; Animals; antibodies; Binding, Competitive; Cell Culture Techniques; chemokines; disease course; Disease Models, Animal; E-Selectin - genetics; E-selectin targeting; endothelial cells; Endothelium; Endothelium, Vascular - drug effects; Endothelium, Vascular - immunology; Female; Gene Expression; gene expression regulation; gene therapy; glomerulonephritis; Glomerulonephritis - genetics; Glomerulonephritis - therapy; HEK293 Cells; Human Umbilical Vein Endothelial Cells; Humans; I-kappa B Proteins - genetics; Inflammation; kidneys; leukocytes; Mice; Mice, Inbred C57BL; NF-kappa B - antagonists & inhibitors; NF-κB inhibition; polyethylene glycol; Protein Binding; signal transduction; Signal Transduction - genetics; transcription factor NF-kappa B; Transgenes; Vascular Cell Adhesion Molecule-1 - genetics; viruses
• ISSN: 0168-3659; 1873-4995
• DOI: 10.1016/j.jconrel.2012.12.016

In chronic inflammatory diseases the endothelium expresses mediators responsible for harmful leukocyte infiltration. We investigated whether targeted delivery of a therapeutic transgene that inhibits nuclear factor κB signal transduction could silence the proinflammatory activation status of endothelial cells. For this, an adenovirus encoding dominant-negative IκB (dnIκB) as a therapeutic transgene was employed. Selectivity for the endothelial cells was achieved by introduction of antibodies specific for inflammatory endothelial adhesion molecules E-selectin or VCAM-1 chemically linked to the virus via polyethylene glycol. In vitro, the retargeted adenoviruses selectively infected cytokine-activated endothelial cells to express functional transgene. The comparison of transductional capacity of both retargeted viruses revealed that E-selectin based transgene delivery exerted superior pharmacological effects. Targeted delivery mediated dnIκB transgene expression in endothelial cells inhibited the induced expression of several inflammatory genes, including adhesion molecules, cytokines, and chemokines. In vivo, in mice suffering from glomerulonephritis, E-selectin-retargeted adenovirus selectively homed in the kidney to microvascular glomerular endothelium. Subsequent downregulation of endothelial adhesion molecule expression 2days after induction of inflammation demonstrated the pharmacological potential of this gene therapy approach. The data justify further studies towards therapeutic virus design and optimization of treatment schedules to investigate their capacity to interfere with inflammatory disease progression. E-selectin retargeted adenovirus encoding dominant negative IkB, homes to inflammatory endothelial cells in the kidney resulting in local downregulation of adhesion molecule expression. [Display omitted]