Genetherapy with adenovirus expressing ATF-BPTI hybrid protein inhibits proteolysis by rheumatoid synovial fibroblasts
article
In rheumatoid arthritis (RA), irreversible joint damage is the result of degradation of articular structures such as cartilage, bone and tendons. The plasminogen activator (PA) system has been shown to be involved in the proteolytic degradation of cartilage matrix by rheumatoid synovial fibroblasts in vitro. To study the possibilities for therapeutic intervention via inhibition of proteolysis by rheumatoid synovial fibroblasts, we compared the effects of genetherapy with three different recombinant adenovirus constructs encoding: 1) the aminoterminal fragment of urokinase (Ad.ATF), a competitive inhibitor of urokinase binding to the urokinase receptor (uPAR); 2) bovine pancreas trypsin inhibitor (Ad.BPTI), also known as Trasylol, a potent plasmin inhibitor; and 3) a hybrid protein of ATF and BPTI (Ad.ATF-BPTI), a plasmin inhibitor that can bind to the urokinase receptor. Synovial fibroblasts were isolated from RA synovial tissue. The cells were infected for 3h with either of the three virus constructs in a concentration of 10s pfu/ml and cultured on a radiolabeled cartilage matrix, produced by bovine chondrocytes cultured in alginate beads. Transfection with Ad.ATF-BPTI resulted in a decrease of matrix degradation of 40%, whereas transfection with the other virus constructs did not inhibit proteolytic activity significantly (<10%). In conclusion, cell surface uPAR bound plasmin inhibition appears to be an effective way to inhibit proteolysis by rheumatoid fibroblasts in vitro. Local inhibition of pericellular proteolysis may be an interesting target for therapeutic intervention to prevent joint destruction in RA.
TNO Identifier
234780
ISSN
13690191
Source
Fibrinolysis and Proteolysis, 12(SUPPL. 1), pp. 13.
Pages
13
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