Cartilage degradation and invasion by rheumatoid synovial fibroblasts is inhibited by gene transfer of TIMP-1 and TIMP-3

Laan, Wijnand; Quax, Paul H.A.; Seemayer, Christian A.; Huisman, L.G.M.; Pieterman, E; Grimbergen, Jos; Verheijen, J.H.; Breedveld, F. C.; Gay, Renate E.; Gay, Steffen; Huizinga, T.W.J.; Pap, Thomas

doi:10.1038/sj.gt.3301871

Cited by 103 publications

(62 citation statements)

References 30 publications

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“…30 In the present study, gene transfer of ribozymes targeting CL also did not completely block cartilage destruction of RA-SF. Therefore, it appears to be evident that the cartilage destructive pathways are not dependent on one executing pathway, but on different matrix degrading enzymes acting in concert.…”

Section: Discussionmentioning

confidence: 73%

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

et al. 2004

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The present study was undertaken to examine whether ribozymes cleaving specifically cathepsin L (CL) mRNA are able to decrease the synthesis of CL protease in rheumatoid arthritis synovial fibroblasts (RA-SF) and thereby reduce the invasiveness into cartilage both in vitro and in the SCID mouse coimplantation model of RA. Two different ribozymes that cleave CL mRNA specifically at positions 533 (RzCL533) and 790 (RzCL790) were generated. Using retroviral gene transfer, RA-SF were transduced with the ribozyme constructs or the empty vector. To examine the effect of the ribozymes on the mRNA level, quantitative analysis for CL mRNA was performed using real-time PCR. For evaluation on the protein level, ELISA using specific anti-CL antibodies was performed. In addition, transduced RA-SF were examined in vitro in a three-dimensional destruction assay evaluating their ability to degrade extracellular matrix produced by human chondrocytes. Matrix destruction was monitored by the release of soluble glycosaminoglycans (sGAG). Using the in vivo SCID mouse coimplantation model of RA, RzCL533-transduced RA-SF and control cells were coimplanted with human cartilage for 60 days. After being killed, invasion of RA-SF into the cartilage was evaluated by using a semiquantitative score. Transduction of RA-SF with RzCL533 and RzCL790 ribozymes decreased significantly the expression of CL mRNA to 44% (range 25-62%) and 20% (range 1-43%), respectively, when compared to mocktransduced cells. The protein concentration of CL in the cell culture supernatants of transduced RA-SF was decreased from 16.0 ng/ml in the mock constructs to 4.1 and 8.2 ng/ml (mean), respectively. Using the in vitro cartilage destruction assay, the release of sGAG decreased to 46 and 60%, respectively, after 14 days when compared to mocktransduced cells. In the SCID mouse coimplantation model of RA, RzCL533-transduced RA-SF revealed a significant lower cartilage invasion when compared to mock and untransduced cells. Using retroviral gene transfer, ribozymes cleaving CL mRNA inhibit specifically the synthesis of this matrix-degrading enzyme and reduce cartilage destruction in in vitro and in vivo models. Our study therefore suggests that ribozymes targeting CL could be a novel and efficient tool to inhibit joint destruction in RA.

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Section: Discussionmentioning

confidence: 73%

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

et al. 2004

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“…It induces apoptosis 2 , inhibits angiogenesis 3 and impedes cell migration 4 , at least in tissue culture or when overexpressed in vivo. Whether these effects occur physiologically is not known.…”

Section: Roy a Blackmentioning

confidence: 99%

“…Moreover, if a deficiency of TIMP3 is detrimental, could elevating its levels be beneficial in inflammatory conditions? Adenoviral delivery to rheumatoid synoviocytes has already been tested 4 , and at least one antiarthritic agent increases TIMP3 expression 15 . The work by Mohammed et al 5 should stimulate many more such investigations.…”

Section: Insights Into Inflammationmentioning

confidence: 99%

“…A series of ten dominant dark skin (Dsk) mutations recently described by Greg Barsh and colleagues 3 can be classified into two categories, depending on whether dermal or epidermal pigmentation is increased. On page 961 of this issue, Van Raamsdonk et al 4 report that three of the four Dsk mutants with increased dermal pigmentation carry activating mutations in Gnaq and Gna11. These genes encode functionally related Gα subunits that mediate the first step in transducing signals from G protein-coupled receptors, identifying a key role for these genes in regulating skin color.…”

Section: The G-netics Of Dark Skinmentioning

confidence: 99%

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TIMP3 checks inflammation

Black

2004

Nat Genet

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“…For gene transfer of TIMP-3, a nonviral expression construct (CMV promoter) as well as replication-defective adenoviruses (E1 deleted, CMV promotor) encoding TIMP-3 (AdTIMP-3) were used (24). The respective constructs without insert (pCMV and AdCMV) as well as untransfected cells served as controls.…”

Section: Gene Transfer Of Timp-3mentioning

confidence: 99%

Gene Transfer of Tissue Inhibitor of Metalloproteinases-3 Reverses the Inhibitory Effects of TNF-α on Fas-Induced Apoptosis in Rheumatoid Arthritis Synovial Fibroblasts

Drynda

Quax

Neumann³

et al. 2005

The Journal of Immunology

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Apart from counteracting matrix metalloproteinases, tissue inhibitor of metalloproteinases-3 (TIMP-3) has proapoptotic properties. These features have been attributed to the inhibition of metalloproteinases involved in the shedding of cell surface receptors such as the TNFR. However, little is known about effects of TIMP-3 in cells that are not susceptible to apoptosis by TNF-α. In this study, we report that gene transfer of TIMP-3 into human rheumatoid arthritis synovial fibroblasts and MRC-5 human fetal lung fibroblasts facilitates apoptosis and completely reverses the apoptosis-inhibiting effects of TNF-α. Although TNF-α inhibits Fas/CD95-induced apoptosis in untransfected and mock-transfected cells, fibroblasts ectopically expressing TIMP-3 are sensitized most strongly to Fas/CD95-mediated cell death by TNF-α. Neither synthetic MMP inhibitors nor glycosylated bioactive TIMP-3 are able to achieve these effects. Gene transfer of TIMP-3 inhibits the TNF-α-induced activation of NF-κB in rheumatoid arthritis synovial fibroblasts and reduces the up-regulation of soluble Fas/CD95 by TNF-α, but has no effects on the cell surface expression of Fas. Collectively, our data demonstrate that intracellularly produced TIMP-3 not only induces apoptosis, but also modulates the apoptosis-inhibiting effects of TNF-α in human rheumatoid arthritis synovial fibroblast-like cells. Thus, our findings may stimulate further studies on the therapeutic potential of gene transfer strategies with TIMP-3.

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Cartilage degradation and invasion by rheumatoid synovial fibroblasts is inhibited by gene transfer of TIMP-1 and TIMP-3

Cited by 103 publications

References 30 publications

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

Targeting cathepsin L (CL) by specific ribozymes decreases CL protein synthesis and cartilage destruction in rheumatoid arthritis

TIMP3 checks inflammation

Gene Transfer of Tissue Inhibitor of Metalloproteinases-3 Reverses the Inhibitory Effects of TNF-α on Fas-Induced Apoptosis in Rheumatoid Arthritis Synovial Fibroblasts

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