Transcription factor decoy for NFkappaB inhibits cytokine and adhesion molecule expressions in synovial cells derived from rheumatoid arthritis.
OBJECTIVE
Numerous cytokines are expressed in lesions of synovial hyperplasia of patients with rheumatoid arthritis (RA), and their pathophysiological contributions have been the subject of speculation. These genes are regulated by the transcription factor NFkappaB which in turn is activated by tumour necrosis factor-alpha (TNF-alpha) and cytokines. In this study we examined the inhibition of the production of pro-inflammatory cytokines, adhesion molecule and matrix metalloproteinase (MMP) from synovial tissue of patients with RA by the introduction of synthetic double-stranded DNA with high affinity for the NFkappaB binding site.


METHODS
NFkappaB decoy oligonucleotides (ODN) were introduced with the aid of the haemagglutinating virus of Japan (HVJ)-liposome method into synovial tissue or synovial cells derived from patients with RA. The levels of interleukin-1beta (IL-1beta), IL-6, TNF-alpha, intercellular adhesion molecule-1 (ICAM-1) and MMP-1 were determined by means of enzyme-linked immunosorbent assay (ELISA) and Northern blotting analysis. A cell counting kit was used to study the effect of NFkappaB decoy ODN on synovial cell proliferation.


RESULTS
The production of these mediators was significantly inhibited by the introduction of NFkappaB decoy ODN compared with the effect of scrambled decoy ODN. Transfection of NFkappaB decoy ODN resulted in a significant inhibition of synovial cell proliferation as compared with that of scrambled decoy ODN.


CONCLUSIONS
The results demonstrated in this study suggest the potential usefulness of NFkappaB decoy ODN for gene therapy of inflammatory synovitis of RA.
Tomita
T
T
Department of Orthopaedic Surgery, Department of Geriatric Medicine, Department of Gene Therapy Science, Osaka University Medical School, 2-2 Yamada-oka, Suita 565-0871, Japan.
Takano
H
H
Tomita
N
N
Morishita
R
R
Kaneko
M
M
Shi
K
K
Takahi
K
K
Nakase
T
T
Kaneda
Y
Y
Yoshikawa
H
H
Ochi
T
T
D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D015815:Cell Adhesion Molecules / Q000096:biosynthesis*; D002455:Cell Division; D016207:Cytokines / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism* / Q000502:physiology; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology; D013583:Synovial Membrane / Q000378:metabolism* / Q000473:pathology; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D015815:Cell Adhesion Molecules; D016207:Cytokines; D007375:Interleukin-1; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D020781:Matrix Metalloproteinase 1; D016650:Fluorescein-5-isothiocyanate