Retroviral gene transfer of an antisense construct against membrane type 1 matrix metalloproteinase reduces the invasiveness of rheumatoid arthritis synovial fibroblasts
Version of Record online: 28 JUN 2005
Copyright © 2005 by the American College of Rheumatology
Arthritis & Rheumatism
Volume 52, Issue 7, pages 2010–2014, July 2005
How to Cite
Rutkauskaite, E., Volkmer, D., Shigeyama, Y., Schedel, J., Pap, G., Müller-Ladner, U., Meinecke, I., Alexander, D., Gay, R. E., Drynda, S., Neumann, W., Michel, B. A., Aicher, W. K., Gay, S. and Pap, T. (2005), Retroviral gene transfer of an antisense construct against membrane type 1 matrix metalloproteinase reduces the invasiveness of rheumatoid arthritis synovial fibroblasts. Arthritis & Rheumatism, 52: 2010–2014. doi: 10.1002/art.21156
- Issue online: 28 JUN 2005
- Version of Record online: 28 JUN 2005
- Manuscript Accepted: 14 APR 2005
- Manuscript Received: 16 JUL 2004
- Swiss National Science Foundation. Grant Number: 32-64142.00
- DFG. Grant Numbers: Pa 698/1-1, Pa 698/3-1, Mu 1383/3-3, Ai 16/10-1
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.
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.
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).
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.