Key result
Transduction of rheumatoid arthritis synovial fibroblasts with MMP-1-specific ribozymes significantly decreased MMP-1 production and reduced cartilage invasion in a SCID mouse model.
Why the study?
Does retroviral gene transfer of MMP-1-specific ribozymes reduce MMP-1 production and invasiveness in rheumatoid arthritis synovial fibroblasts?
Does retroviral gene transfer of MMP-1-specific ribozymes reduce MMP-1 production and invasiveness in rheumatoid arthritis synovial fibroblasts?
Intracellular expression of MMP-1-specific ribozymes significantly reduces MMP-1 production and cartilage invasion by rheumatoid arthritis synovial fibroblasts in a SCID mouse model.
Supports MMP-1 ribozymes as potential RA target; leaves open translation from SCID mouse models to human therapy.
OBJECTIVE: To investigate whether retroviral gene transfer of ribozymes targeting matrix metalloproteinase 1 (MMP-1) inhibits the production of MMP-1 in rheumatoid arthritis synovial fibroblasts (RASFs) and reduces the invasiveness of these cells in vivo. METHODS: MMP-1-specific ribozymes (RzMMP-1) were designed and cloned into the pLNSX retroviral vector. Cleavage of MMP-1 was determined in vitro, and the most effective ribozyme was selected for further investigation. RASFs were transduced with replication-deficient viruses carrying RzMMP-1 or with empty viruses (mock). Quantitative polymerase chain reaction with cleavage site-spanning fluorescent probes was used to measure the levels of MMP-1, MMP-9, and MMP-13 messenger RNA. In addition, protein levels of MMP-1 in cell culture supernatants were determined by enzyme linked immunosorbent assay. The effects of stimulation with lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNFalpha) on the production of MMP-1 were assessed accordingly. The invasiveness of RzMMP-1-transduced, mock-transduced, and untransduced RASFs was analyzed in the SCID mouse in vivo model of RA. RESULTS: Transduction of RASFs with RzMMP-1 significantly decreased the production of MMP-1 in RASFs without affecting other MMPs, such as MMP-9 and MMP-13. RzMMP-1 not only reduced the spontaneous production of MMP-1, but also prevented the LPS- and TNFalpha-induced increase in MMP-1 production. Inhibition of MMP-1 was maintained for at least 2 months and was accompanied by a significant reduction of the invasiveness of RASFs in the SCID mouse model of RA. CONCLUSION: Intracellular expression of ribozymes constitutes a feasible tool for inhibiting the production of matrix-degrading enzymes. Inhibition of MMP-1 alone results in a significant reduction of cartilage invasion by RASFs.
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Rutkauskaite et al. (2004) studied Rheumatoid arthritis. Retroviral gene transfer of MMP-1-specific ribozymes (RzMMP-1) vs. Empty viruses (mock) or untransduced cells was evaluated on Production of MMP-1 and invasiveness of RASFs in the SCID mouse model. Transduction of rheumatoid arthritis synovial fibroblasts with MMP-1-specific ribozymes significantly decreased MMP-1 production and reduced cartilage invasion in a SCID mouse model.
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