title: Citrullination of fibronectin modulates synovial fibroblast behavior.
abstract: BACKGROUND
  Rheumatoid arthritis is an autoimmune arthritis characterized by joint destruction. Anti-citrullinated protein antibodies are pathologic in rheumatoid arthritis, but the role of the citrullinated proteins themselves is much less clear. Citrullination is the conversion of the arginine residues of a protein to citrulline. In the inflamed rheumatoid joint there is increased protein citrullination. Several proteins are citrullinated in rheumatoid arthritis, including collagen type II, fibrinogen, and fibronectin. Fibronectin is thought to mediate the adhesion of joint-invading synovial fibroblasts to the rheumatoid cartilage in addition to regulating other synovial fibroblast functions. However, the effect of citrullinated fibronectin on synovial fibroblasts is unknown.
  
  
  METHODS
  To investigate the effect of citrullinated fibronectin on synovial fibroblast behavior, we cultured normal murine, arthritic murine, and human rheumatoid synovial fibroblasts. We then compared several synovial fibroblast functions in the presence of fibronectin versus citrullinated fibronectin. We assessed adhesion with time-lapse microscopy, migration with transwell assays, focal adhesion kinase and paxillin phosphorylation by western blot, and focal matrix degradation by fluorescent gelatin degradation.
  
  
  RESULTS
  Normal synovial fibroblasts have impaired adhesion, spreading, migration, and integrin-mediated phosphorylation of focal adhesion kinase and paxillin on citrullinated fibronectin. Murine arthritic and human rheumatoid synovial fibroblasts also have impaired adhesion and spreading on citrullinated fibronectin, but focal matrix degradation is unaffected by citrullinated fibronectin.
  
  
  CONCLUSIONS
  Citrullination of fibronectin alters synovial fibroblast behavior and may affect how these cells adhere to and invade the joint and travel through the bloodstream. This work suggests an important role for the interaction of synovial fibroblasts with citrullinated matrix in the pathophysiology of rheumatoid arthritis.
authors.forename: Miriam A
authors.initials: MA
authors.lastname: Shelef
authors.forename: David A
authors.initials: DA
authors.lastname: Bennin
authors.forename: Deane F
authors.initials: DF
authors.lastname: Mosher
authors.forename: Anna
authors.initials: A
authors.lastname: Huttenlocher
mesh_terms: D000818:Animals; D000843:Ankle Joint / Q000166:cytology; D001168:Arthritis / Q000378:metabolism / Q000473:pathology; D001172:Arthritis, Rheumatoid / Q000378:metabolism / Q000473:pathology; D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D002478:Cells, Cultured; D002956:Citrulline / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D051419:Paxillin / Q000378:metabolism; D010766:Phosphorylation; D013582:Synovial Fluid / Q000166:cytology
chemical_list: D005353:Fibronectins; D016023:Integrins; D051419:Paxillin; D002956:Citrulline; D005780:Gelatin; D051416:Focal Adhesion Protein-Tyrosine Kinases