Therapeutic effect of in vivo transfection of transcription factor decoy to NF-κB on septic lung in mice

Abstract

Nuclear factor-kappaB (NF-kappaB) plays a key role in regulating expression of several genes involved in the pathophysiology of endotoxic shock. We investigated whether in vivo introduction of synthetic double-stranded DNA with high affinity for the NF-kappaB binding site could block expression of genes mediating pulmonary vascular permeation and thereby provide effective therapy for septic lung failure. Endotoxic shock was induced by an intravenous injection of 10 mg/kg Escherichia coli endotoxin in mice. We introduced NF-kappaB decoy oligodeoxynucleotide (ODN) in vivo 1 h after endotoxic shock by using a gene transfer kit. At 10 h, blood samples were collected for measurement of histamine and for blood-gas analysis. Gene and protein expression levels of target molecules were determined by means of Northern and Western blot analyses, respectively. The transpulmonary flux of (125)I-labeled albumin was used as an index of lung vascular permeability. Administration of endotoxin caused marked increases in plasma histamine and gene and protein expressions of histidine decarboxylase, histamine H(1) receptors, and inducible nitric oxide synthase in lung tissues. Elevated lung vascular permeability was also found. Blood-gas analysis showed concurrent decreases in arterial Po(2), Pco(2), and pH. All of these events induced by endotoxin were significantly inhibited by transfection of NF-kappaB decoy ODN but not by its mutated (scrambled) form (used as a control). Our results indicate for the first time the potential usefulness of NF-kappaB decoy ODN for gene therapy of endotoxic shock.

Vascular biology in sepsis: pathophysiological and therapeutic significance of vascular dysfunction

“…We have shown that eNOS expression is obviously diminished in blood vessels from rabbits (Matsuda et al, 2003) and in lung tissues from mice (Matsuda et al, 2004) following induction of sepsis with LPS. In our recent work, furthermore, it has been demonstrated that sepsis causes a progressive and profound reduction in phosphorylation of eNOS in rabbit mesenteric arteries (Matsuda et al, 2006), suggesting less production of NO by eNOS in sepsis.…”

Gene therapy progress and prospects: RNA aptamers

“…The decoy has been tested in several arthritis models as well as in endotoxin-induced models of liver failure and septic lung in mice. [13][14][15] Currently, Avrina is in Phase I/II trials for treatment of mild-to-moderate atopic dermatitis. In addition, a phosphorothioated NF-kB decoy was recently tested in a murine inflammatory bowel disease model.…”

Decoy oligodeoxynucleotide targeting activator protein-1 (AP-1) attenuates intestinal inflammation in murine experimental colitis

“…Treatment with AP-1 or NF-kB decoy ODN significantly decreased IL-1b, TNF-a, and MPO activities in the tissue contents, which coincided well with the results of our histological examinations (Figure 6a-c). [33][34][35][36][37][38][39][40] Although AP-1 is a major immunoregulatory as well as proinflammatory transcription factor, 21,[27][28][29][30][45][46][47] nothing is known to have been reported regarding the effect of AP-1 inhibition by a decoy ODN on intestinal inflammation. In the present study, we designed a ds decoy ODN binding to AP-1-specific nucleotide sequences and investigated its efficacy to prevent murine experimental DSS-induced colitis.…”

“…33,34 The multiplicity of target genes that are under the control of a single transcription factor also contributes to utility of decoy ODN strategies. Recent studies have demonstrated therapeutic applications of several decoy ODNs targeting transcription factors including NF-kB, [35][36][37] AP-1, 38-40 E2F, 54 Sp1, 55,56 and the signal transducer and activator of transcription (STAT) family. 57,58 There have also been a few reports suggesting an active role of AP-1 during the pathogenesis of IBD.…”

“…33,34 It has been reported that a decoy ODN targeted at the transcription factor nuclear factor (NF)-kB, which plays an essential role in the regulation of a large number of genes involved in immune and inflammatory response, can exert potent immunosuppressive effects with certain inflammatory diseases. [35][36][37] Although the effects of AP-1 decoy ODN have been recently studied in several experimental disease models, [38][39][40] nothing is known regarding its therapeutic potential in gastrointestinal inflammation, including IBD.…”