Tgf Beta Responsive Tyrosine Phosphatase Promotes Rheumatoid Synovial Fibroblast Invasiveness

Objective In rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) that line joint synovial membranes aggressively invade the extracellular matrix, destroying cartilage and bone. As signal transduction in FLS is mediated through multiple pathways involving protein tyrosine phosphorylation, we sought to identify protein tyrosine phosphatases (PTPs) regulating the invasiveness of RA FLS. We describe that the transmembrane receptor PTP. (RPTP.), encoded by the transforming growth factor (TGF) beta-target gene, PTPRK, promotes RA FLS invasiveness.Methods Gene expression was quantified by quantitative PCR. PTP knockdown was achieved using antisense oligonucleotides. FLS invasion and migration were assessed in transwell or spot assays. FLS spreading was assessed by immunofluorescence microscopy. Activation of signalling pathways was analysed by Western blotting of FLS lysates using phosphospecific antibodies. In vivo FLS invasiveness was assessed by intradermal implantation of FLS into nude mice. The RPTP. substrate was identified by pull-down assays.Results PTPRK expression was higher in FLS from patients with RA versus patients with osteoarthritis, resulting from increased TGFB1 expression in RA FLS. RPTP. knockdown impaired RA FLS spreading, migration, invasiveness and responsiveness to platelet-derived growth factor, tumour necrosis factor and interleukin 1 stimulation. Furthermore, RPTP. deficiency impaired the in vivo invasiveness of RA FLS. Molecular analysis revealed that RPTP. promoted RA FLS migration by dephosphorylation of the inhibitory residue Y527 of SRC.Conclusions By regulating phosphorylation of SRC, RPTP. promotes the pathogenic action of RA FLS, mediating cross-activation of growth factor and inflammatory cytokine signalling by TGF beta in RA FLS.