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Open Access Highly Accessed Research article

Citrullination of fibronectin modulates synovial fibroblast behavior

Miriam A Shelef1*, David A Bennin23, Deane F Mosher14 and Anna Huttenlocher23

Author Affiliations

1 Department of Medicine, Division of Rheumatology, University of Wisconsin - Madison, 1685 Highland Avenue, Madison, WI 53705, USA

2 Department of Pediatrics, University of Wisconsin - Madison, 1550 Linden Drive, Madison, WI 53706, USA

3 Department of Medical Microbiology and Immunology, University of Wisconsin - Madison, 1550 Linden Drive, Madison, WI 53706, USA

4 Department of Biomolecular Chemistry, University of Wisconsin - Madison, 1300 University Avenue, Madison, WI 53706, USA

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Arthritis Research & Therapy 2012, 14:R240  doi:10.1186/ar4083

Published: 5 November 2012

Abstract

Introduction

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.

Conclusion

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.