Constitutive upregulation of the transforming growth factor-beta pathway in rheumatoid arthritis synovial fibroblasts.

Abstract

Genome-wide gene expression was comparatively investigated in early-passage rheumatoid arthritis (RA) and osteoarthritis (OA) synovial fibroblasts (SFBs; n = 6 each) using oligonucleotide microarrays; mRNA/protein data were validated by quantitative PCR (qPCR) and western blotting and immunohistochemistry, respectively. Gene set enrichment analysis (GSEA) of the microarray data suggested constitutive upregulation of components of the transforming growth factor (TGF)-beta pathway in RA SFBs, with 2 hits in the top 30 regulated pathways. The growth factor TGF-beta1, its receptor TGFBR1, the TGF-beta binding proteins LTBP1/2, the TGF-beta-releasing thrombospondin 1 (THBS1), the negative effector SkiL, and the smad-associated molecule SARA were upregulated in RA SFBs compared to OA SFBs, whereas TGF-beta2 was downregulated. Upregulation of TGF-beta1 and THBS1 mRNA (both positively correlated with clinical markers of disease activity/severity) and downregulation of TGF-beta2 mRNA in RA SFBs were confirmed by qPCR. TGFBR1 mRNA (only numerically upregulated in RA SFBs) and SkiL mRNA were not differentially expressed. At the protein level, TGF-beta1 showed a slightly higher expression, and the signal-transducing TGFBR1 and the TGF-beta-activating THBS1 a significantly higher expression in RA SFBs than in OA SFBs. Consistent with the upregulated TGF-beta pathway in RA SFBs, stimulation with TGF-beta1 resulted in a significantly enhanced expression of matrix-metalloproteinase (MMP)-11 mRNA and protein in RA SFBs, but not in OA SFBs. In conclusion, RA SFBs show broad, constitutive alterations of the TGF-beta pathway. The abundance of TGF-beta, in conjunction with an augmented mRNA and/or protein expression of TGF-beta-releasing THBS1 and TGFBR1, suggests a pathogenetic role of TGF-beta-induced effects on SFBs in RA, for example, the augmentation of MMP-mediated matrix degradation/remodeling.

• Authors: Dirk Pohlers, Andreas Beyer, Dirk Koczan, Thomas Wilhelm, Hans-Jürgen Thiesen, Raimund W Kinne
• Journal: Arthritis research & therapy (Arthritis Res Ther) Vol. 9 Issue 3 Pg. R59 ( 2007) ISSN: 1478-6362 [Electronic] England
• PMID: 17594488 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Messenger
• Receptors, Transforming Growth Factor beta
• Thrombospondin 1
• Transforming Growth Factor beta1

Topics

• Aged
• Arthritis, Rheumatoid (genetics, metabolism)
• Blotting, Western
• Female
• Fibroblasts (metabolism)
• Gene Expression
• Gene Expression Profiling
• Humans
• Immunohistochemistry
• Male
• Middle Aged
• Oligonucleotide Array Sequence Analysis
• RNA, Messenger (analysis)
• Receptors, Transforming Growth Factor beta (metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• Synovial Membrane (cytology, metabolism)
• Thrombospondin 1 (metabolism)
• Transforming Growth Factor beta1 (metabolism)
• Up-Regulation