Retroviral gene transfer of an antisense construct against membrane type 1 matrix metalloproteinase reduces the invasiveness of rheumatoid arthritis synovial fibroblasts

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Abstract
Objective Membrane type 1 matrix metalloproteinase (MT1‐MMP) is expressed prominently in rheumatoid arthritis synovial fibroblasts (RASFs), but the specific contribution of MT1‐MMP to fibroblast‐mediated destruction of articular cartilage is incompletely understood. This study used gene transfer of an antisense expression construct to assess the effects of MT1‐MMP inhibition on the invasiveness of RASFs. Methods Retroviral gene transfer of a pLXIN vector–based antisense RNA expression construct (MT1‐MMPαS) to MT1‐MMP was used to stably transduce RASFs. Levels of MT1‐MMP RNA and protein were determined by quantitative polymerase chain reaction, Western blotting, and immunocytochemistry in MT1‐MMPαS–transduced RASFs as well as in control cells, with monitoring for 60 days. The effects of MT1‐MMPαS on the invasiveness of RASFs were analyzed in the SCID mouse coimplantation model of RA. Results MT1‐MMPαS–transduced RASFs produced high levels of antisense RNA that exceeded endogenous levels of MT1‐MMP messenger RNA by 15‐fold and resulted in a down‐regulation of MT1‐MMP at the protein level. Inhibition of MT1‐MMP production was maintained for 60 days and significantly reduced the invasiveness of RASFs in the SCID mouse model. Whereas prominent invasion into cartilage by nontransduced and mock‐transduced RASFs was observed (mean invasion scores 3.0 and 3.1, respectively), MT1‐MMPαS–transduced cells showed only moderate invasiveness (mean invasion score 1.8; P < 0.05). Conclusion The data demonstrate that an antisense RNA expression construct against MT1‐MMP can be generated and expressed in RASFs for at least 60 days. Inhibition of MT1‐MMP significantly reduces the cartilage degradation by RASFs.

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