Transcription factor decoy for NFκB inhibits TNF-α-induced cytokine and adhesion molecule expression in vivo

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Abstract
The expression of several cytokines and adhesion molecules is regulated by the transcription factor NFκB, which is activated by tumor necrosis factor alpha (TNF-α). In this study, we employed a mouse model of nephritis induced by TNF-α to examine whether inhibition of NFκB activity using transcription factor decoy oligonucleotides (ODN) blocks cytokine and adhesion molecule expression and attenuates the renal inflammatory response. First, we developed a method for delivering FITC-ODN in vivo into mouse kidney glomeruli by employing HVJ-liposome. Then, in order to study the feasibility of decoy strategy in vivo, the reporter gene chloramphenicol acetyltransferase (CAT) driven by three tandemly repeated NFκB binding sequences was transfected into the kidney. Intrapenetorial injection of TNF-α stimulated CAT expression in vivo, and the increase in CAT expression was completely abolished by NFκB decoy ODN, but not scrambled ODN. Therefore, we examined the effect of NFκB decoy ODN transfection on TNF-α-induced endogenous interleukin (IL)-1α, IL-1β, IL-6, ICAM-1 and VCAM-1 gene expression as assessed by RT-PCR and Northern blotting. Our present data showed that NFκB decoy, but not scrambled, ODN abolished TNF-α induced gene expression in vivo, as well as glomerular inflammation as assessed by CD45 staining. Taken together, our results suggest the potential utility of NFκB decoy strategy for molecular therapy to glomerular inflammatory diseases.