Functional inhibition of NF-κB signal transduction in αvβ3 integrin expressing endothelial cells by using RGD-PEG-modified adenovirus with a mutant IκB gene

Abstract

In order to selectively block nuclear factor κB (NF-κB)- dependent signal transduction in angiogenic endothelial cells, we constructed an αvβ3 integrin specific adenovirus encoding dominant negative IκB (dnIκB) as a therapeutic gene. By virtue of RGD modification of the PEGylated virus, the specificity of the cell entry pathway of adenovirus shifted from coxsackiadenovirus receptor dependent to αvβ3 integrin dependent entry. The therapeutic outcome of delivery of the transgene into endothelial cells was determined by analysis of cellular responsiveness to tumor necrosis factor (TNF)-α. Using real time reverse transcription PCR, mRNA levels of the cell adhesion molecules E-selectin, vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1, the cytokines/growth factors IL-6, IL-8 and vascular endothelial growth factor (VEGF)-A, and the receptor tyrosine kinase Tie-2 were assessed. Furthermore, levels of ICAM-1 protein were determined by flow cytometric analysis. RGD-targeted adenovirus delivered the dnIκB via αvβ3 to become functionally expressed, leading to complete abolishment of TNF-α-induced up-regulation of E-selectin, ICAM-1, VCAM-1, IL-6, IL-8, VEGFA and Tie-2. The approach of targeted delivery of dnIκB into endothelial cells presented here can be employed for diseases such as rheumatoid arthritis and inflammatory bowel disease where activation of NF-κB activity should be locally restored to basal levels in the endothelium. © 2006 Fishman et al, licensee BioMed Central Ltd.