title: Amelioration of arthritis by intraarticular dominant negative Ikk beta gene therapy using adeno-associated virus type 5.
abstract: Nuclear factor (NF)-kappaB is highly activated in the synovium of rheumatoid arthritis (RA) patients, and can induce transcription of many proinflammatory molecules. Phosphorylation of inhibitor of kappaB (IkappaB) proteins is an important step in NF-kappaB activation and under inflammatory conditions is regulated predominantly by IkappaB kinase (IKK)beta. Consequently, specific targeting of IKK beta in the joint, using gene therapy, presents a sophisticated treatment option for arthritis. In the present study we investigated the effect of inhibiting IKK beta in adjuvant arthritis (AA) in rats, using recombinant adeno-associated virus (rAAV)-mediated intraarticular gene therapy. For this purpose rAAV5 carrying the dominant negative IKK beta gene (AAV5.IKK beta dn) or control AAV5.eGFP was injected into the right ankle joint. Rats treated with AAV5.IKK beta dn in early arthritis exhibited significantly reduced paw swelling (p < 0.05). Immunohistochemical analysis of synovial tissue revealed reduced levels of interleukin (IL)-6 (p = 0.005) and tumor necrosis factor-alpha (TNF-alpha) (p = 0.03), whereas IL-10 levels were not affected. No significant effect was found on cartilage and bone destruction, or on matrix metalloproteinase-3 and tissue inhibitor of matrix metalloproteinase-1 expression. Injection of AAV5.IKK beta dn in the preclinical phase showed only a marginal effect on arthritis. Importantly, in this study we also demonstrate for the first time that our vector is capable of transducing human RA whole synovial tissue biopsies ex vivo, resulting in reduced IL-6 production after TNF-alpha stimulation (p = 0.03). In conclusion, we are the first to demonstrate that rAAV5 can be used to successfully deliver a therapeutic gene (IKK beta dn) to the synovium, resulting in reduced severity of inflammation in AA in vivo and proinflammatory cytokine production in human RA synovial tissue ex vivo. This translational research represents a crucial next step toward the development of gene therapy for application in humans.
authors.affiliation: Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, 1100 DE Amsterdam, The Netherlands.
authors.forename: Sander W
authors.initials: SW
authors.lastname: Tas
authors.forename: Janik
authors.initials: J
authors.lastname: Adriaansen
authors.forename: Najat
authors.initials: N
authors.lastname: Hajji
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Bakker
authors.forename: Gary S
authors.initials: GS
authors.lastname: Firestein
authors.forename: Margriet J
authors.initials: MJ
authors.lastname: Vervoordeldonk
authors.forename: Paul P
authors.initials: PP
authors.lastname: Tak
mesh_terms: D000818:Animals; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus; D005799:Genes, Dominant; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics* / Q000378:metabolism; D020672:I-kappa B Proteins / Q000235:genetics / Q000378:metabolism; D007249:Inflammation / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D008297:Male; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D011917:Rats, Inbred Lew; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology
chemical_list: D016207:Cytokines; D020672:I-kappa B Proteins; D016328:NF-kappa B; D051550:I-kappa B Kinase