Suppression of the inflammatory response by disease-inducible interleukin-10 gene therapy in a three-dimensional micromass model of the human synovial membrane.
BACKGROUND
Gene therapy has the potential to provide long-term production of therapeutic proteins in the joints of osteoarthritis (OA) patients. The objective of this study was to analyse the therapeutic potential of disease-inducible expression of anti-inflammatory interleukin-10 (IL-10) in the three-dimensional micromass model of the human synovial membrane.


METHODS
Synovial tissue samples from OA patients were digested and the cells were mixed with Matrigel to obtain 3D micromasses. The CXCL10 promoter combined with the firefly luciferase reporter in a lentiviral vector was used to determine the response of the CXCL10 promoter to tumour necrosis factor alpha (TNF-α), interleukin-1β (IL-1β) and lipopolysaccharide (LPS). The effects of recombinant IL-10 on gene expression were determined by quantitative PCR. The production of IL-10 from the CXCL10p-IL10 vector and the effects on pro-inflammatory cytokine production were assessed by multiplex ELISA.


RESULTS
Micromasses made from whole synovial membrane cell suspensions form a distinct surface composition containing macrophage and fibroblast-like synoviocytes thus mimicking the synovial lining. This lining can be transduced by lentiviruses and allow CXCL-10 promoter-regulated transgene expression. Adequate amounts of IL-10 transgene were produced after stimulation with pro-inflammatory factors able to reduce the production of synovial IL-1β and IL-6.


CONCLUSIONS
Synovial micromasses are a suitable model to test disease-regulated gene therapy approaches and the CXCL10p-IL10 vector might be a good candidate to decrease the inflammatory response implicated in the pathogenesis of OA.
['Broeren|Mathijs G A|MG|', 'de Vries|Marieke|M|', 'Bennink|Miranda B|MB|', 'Arntz|Onno J|OJ|', 'van Lent|Peter L E M|PL|', 'van der Kraan|Peter M|PM|', 'van den Berg|Wim B|WB|', 'van den Hoogen|Frank H J|FH|', 'Koenders|Marije I|MI|', 'van de Loo|Fons A J|FA|']
D054357:Chemokine CXCL10 D004797:Enzyme-Linked Immunosorbent Assay D005260:Female D005434:Flow Cytometry D015316:Genetic Therapy D005822:Genetic Vectors D006801:Humans D016753:Interleukin-10 D016086:Lentivirus D008297:Male D018613:Microscopy, Confocal D010003:Osteoarthritis D011401:Promoter Regions, Genetic D060888:Real-Time Polymerase Chain Reaction D013583:Synovial Membrane D046509:Tissue Culture Techniques
Gene therapy Micromasses Osteoarthritis Cytokines Inflammation Synovium