Adenoviral gene transfer of tissue inhibitors of metalloproteinases (TIMPs) reduces the invasive behaviour of rheumatoid fibroblast-like synovicytes

In rheumatoid arthritis (RA), an excess of proteolytic enzymes secreted at the synovium-cartilage junction results in the invasion of the articular cartilage by synovial cells. The aims of the present study were to investigate the effects of overexpression of TIMP-1 and TIMP-3 on: 1) the invasive behaviour of rheumatoid fibroblasts-like synoviocytes and 2) cell proliferation and apoptosis. The day before the experiments, the synoviocytes were infected with adenoviral vectors encoding TIMP-1 or TIMP-3 or with a control vector (AdLacZ). A Transwell system was used to study invasion of the cells. After 3 days, the invaded cells were counted using a microscope. Proliferation was assessed by measuring 3H-thymidine incorporation and cell counting. Apoptosis was assessed at 1-4 days after transduction using an Annexin V-FITC kit. Both TIMP-1 and TIMP-3 overexpression resulted in a significant reduction, respectively 60% (P < 0.001) and 80% (P < 0.001), of invasiveness of the synoviocytes as compared to the AdLacZ-transduced cells. In all cases, TIMP-3 was superior to TIMP-1 (P = 0.02). Cell proliferation was significantly reduced by TIMP-3 overexpression (40%; P < 0.05) and to a lesser extend by TIMP-1 (20%; P < 0.05) as compared to AdLacZ. There were little differences in % of apoptotic cells between the non-transduced, AdTIMP-1, AdTIMP-3 or Ad LacZ transduced cells up to 4 days after transduction. A maximum of 15% of the AdTIMP-3 transduced cells were in apoptotis as compared to a maximum of 12% in the other conditions. These results show that the invasive behavior of RA-FLSs can be strongly inhibited by overexpression of TIMPs. Both MMP inhibition and a reduction of proliferation appear to contribute to this effect. The superior effect of TIMP-3 may be due to a stronger effect on proliferation or to differences in the inhibitory profile of TIMP-1 and TIMP-3. To limit joint destruction in rheumatoid arthritis, inhibition of cartilage invasion by the pannus tissue by TIMP overexpression may be a useful approach.