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

The expression of several cytokines and adhesion molecules is regulated by the transcription factor NF kappa B, which is activated by tumor necrosis factor alpha (TNF-alpha). In this study, we employed a mouse model of nephritis induced by TNF-alpha to examine whether inhibition of NF kappa B 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of decoy strategy in vivo, the reporter gene chloramphenicol acetyltransferase (CAT) driven by three tandemly repealed NF kappa B binding sequences was transfected into the kidney. Intrapenetorial injection of TNF-alpha stimulated CAT expression in vivo, and the increase in CAT expression was completely abolished by NF kappa B decoy ODN, but not scrambled ODN. Therefore, we examined the effect of NF kappa B decoy ODN transfection on TNF-alpha-induced endogenous interleukin (IL)-1 alpha, IL-1 beta, IL-6, ICAM-1 and VCAM-1 gene expression as assessed by RT-PCR and Northern blotting. Our present data showed that NF kappa B decoy, but not scrambled, ODN abolished TNF-alpha induced gene expression in vivo, as well as glomerular inflammation as assessed by CD45 staining. Taken together, our results suggest the potential utility of NF kappa B decoy strategy for molecular therapy to glomerular inflammatory diseases.