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 coxsacki-adenovirus 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, VEGF-A 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.
Highlights
Microvascular endothelial cells are active participants in a variety of diseases, including cancer [1] and chronic inflammation such as rheumatoid arthritis [2]
We showed that PEGylation of the adenovirus and subsequent conjugation with anti-E-selectin antibody as a homing ligand coupled onto the distal functional group of polyethylene glycol (PEG) could selectively deliver a reporter gene into activated endothelial cells in vivo
nuclear factor κB (NF-κB) is a transcription factor that controls the expression of cytokines, chemokines and endothelial cell adhesion molecules to facilitate leukocyte movement from the blood stream into the underlying tissue [18,19]
Summary
Microvascular endothelial cells are active participants in a variety of diseases, including cancer [1] and chronic inflammation such as rheumatoid arthritis [2]. Inflammatory mediators can either directly or indirectly, promote angiogenesis. Several observations suggest that angiogenesis and inflammation proceed in a co-ordinated fashion and sustain one another during chronic inflammatory diseases and in cancer growth [4]. Their active roles in the pathophysiology of disease, together with their easy accessibility in the blood, makes endothelial cells attractive target cells for therapy. Nuclear factor κB (NF-κB)/Rel transcription factors represent a ubiquitously expressed protein family that modulates the expression of genes involved in diverse cellular functions, such
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