Citrullination of fibronectin in rheumatoid arthritis synovial tissue

• Published in: Rheumatology (Oxford, England) Vol. 44; no. 11; pp. 1374 - 1382
• Main Authors: Chang, X., Yamada, R., Suzuki, A., Kochi, Y., Sawada, T., Yamamoto, K.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.11.2005; Oxford Publishing Limited (England)
• Subjects: Apoptosis - drug effects; Arthritis, Rheumatoid - metabolism; Biological and medical sciences; Blotting, Western - methods; Cells, Cultured; Citrullination; Citrulline - blood; Citrulline - metabolism; Citrulline - pharmacology; Diseases of the osteoarticular system; Enzyme-Linked Immunosorbent Assay - methods; Fibronectin; Fibronectins - blood; Fibronectins - metabolism; Fibronectins - pharmacology; HL-60 Cells; Humans; Inflammatory joint diseases; Integrin beta1 - metabolism; Integrin β1; Medical sciences; Osteoarthritis - metabolism; PAD4 or PAD5; Peptidylarginine deiminase 4 (PADI4; Rheumatoid arthritis; Synovial Membrane - metabolism; Synovial tissue; Vascular Endothelial Growth Factor A - metabolism; VEGF; Citrullination; Peptidylarginine deiminase 4 PADI4; VEGF; Diseases of the osteoarticular system; Rheumatology; Autoimmune disease; Inflammatory joint disease; Fibronectin; Chronic; Integrin; Rheumatoid arthritis; PAD4 or PADS; Synovial tissue; Integrin β1
• ISSN: 1462-0324; 1462-0332
• DOI: 10.1093/rheumatology/kei023

Objectives. Citrullination, catalysed by peptidylarginine deiminase (PAD), is the post-translational modification of peptidylarginine to citrulline, which is intimately involved in the pathogenesis of rheumatoid arthritis (RA). Fibronectin (Fn), a large glycoprotein, is expressed at high levels in arthritic joints and it mediates various physiological processes through interactions with cell-surface integrin receptors and growth factors. We investigated the citrullination of Fn and its potential contribution to the pathogenesis of RA. Methods. We localized Fn expression and citrullination in RA synovial tissue by immunohistochemistry, immunoprecipitation and western blotting. We also determined levels of citrullinated Fn in plasma from RA patients using sandwich enzyme-linked immunosorbent assay (ELISA). After incubating Fn with rabbit skeletal muscle PAD, we examined the binding ability of citrullinated Fn to vascular endothelial growth factor (VEGF) and integrin β1 using a solid-phase receptor binding assay as well as the effect of the citrullinated Fn on apoptosis using cultured HL-60 cells. Results. Immunohistochemistry and western blotting analysis indicated that Fn formed extracellular aggregates that were specifically citrullinated in RA synovial tissue. No Fn deposits were observed in synovial tissues of osteoarthritis (OA). Sandwich ELISA detected higher levels of citrullinated Fn in plasma from patients with RA than from healthy controls or those with systemic lupus erythematosus. Following citrullination in vitro, the affinity of Fn for VEGF increased, but binding activity to integrin β1 decreased and Fn no longer stimulated the apoptosis of monocytes induced from cultured HL-60 cells. Conclusions. Our results suggest that the citrullination of Fn is a specific event for RA synovium, although others have detected citrullinated total proteins in inflamed synovial tissue of RA and non-RA patients. Citrullination of Fn could alter interactions between Fn and its receptors and growth factors, consequently contributing to mechanisms of RA pathogenesis such as perturbed angiogenesis and apoptosis.