title: Disease-Regulated Gene Therapy with Anti-Inflammatory Interleukin-10 Under the Control of the CXCL10 Promoter for the Treatment of Rheumatoid Arthritis.
abstract: Disease-inducible promoters for the treatment of rheumatoid arthritis (RA) have the potential to provide regulated expression of therapeutic proteins in arthritic joints. In this study, we set out to identify promoters of human genes that are upregulated during RA and are suitable to drive the expression of relevant amounts of anti-inflammatory interleukin (IL)-10. Microarray analysis of RA synovial biopsies compared with healthy controls yielded a list of 22 genes upregulated during RA. Of these genes, CXCL10 showed the highest induction in lipopolysaccharide-stimulated synovial cells. The CXCL10 promoter was obtained from human cDNA and cloned into a lentiviral vector carrying firefly luciferase to determine the promoter inducibility in primary synovial cells and in THP-1 cells. The promoter activation was strongest 8-12 hr after stimulation with the proinflammatory cytokine tumor necrosis factor (TNF)-α and was reinducible after 96 hr. In addition, the CXCL10 promoter showed a significant response to RA patient serum, compared with sera from healthy individuals. The luciferase gene was replaced with IL-10 to determine the therapeutic properties of the CXCL10p-IL10 lentiviral vector. Primary synovial cells transduced with CXCL10p-IL10 showed a great increase in IL-10 production after stimulation, which reduced the release of proinflammatory cytokines TNF-α and IL-1β. We conclude that the selected proximal promoter of the CXCL10 gene responds to inflammatory mediators present in the serum of patients with RA and that transduction with the lentiviral CXCL10p-IL10 vector reduces inflammatory cytokine production by primary synovial cells from patients with RA. CXCL10 promoter-regulated IL-10 overexpression can thus provide disease-inducible local gene therapy suitable for RA.
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Mathijs G A
authors.initials: MG
authors.lastname: Broeren
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Marieke
authors.initials: M
authors.lastname: de Vries
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Miranda B
authors.initials: MB
authors.lastname: Bennink
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Onno J
authors.initials: OJ
authors.lastname: Arntz
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Arjen B
authors.initials: AB
authors.lastname: Blom
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Marije I
authors.initials: MI
authors.lastname: Koenders
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Peter L E M
authors.initials: PL
authors.lastname: van Lent
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Peter M
authors.initials: PM
authors.lastname: van der Kraan
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Wim B
authors.initials: WB
authors.lastname: van den Berg
authors.affiliation: Experimental Rheumatology, Radboud University Medical Center , Nijmegen, The Netherlands .
authors.forename: Fons A J
authors.initials: FA
authors.lastname: van de Loo
mesh_terms: D001172:Arthritis, Rheumatoid / Q000235:genetics* / Q000378:metabolism / Q000628:therapy; D002460:Cell Line; D054357:Chemokine CXCL10 / Q000235:genetics*; D016207:Cytokines / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D011401:Promoter Regions, Genetic; D013582:Synovial Fluid / Q000378:metabolism; D019076:Transgenes
chemical_list: D054357:Chemokine CXCL10; D016207:Cytokines; D016753:Interleukin-10