Differential proteomic analysis of synovial fluid from rheumatoid arthritis and osteoarthritis patients

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Abstract

Background

Rheumatoid arthritis and osteoarthritis are two common musculoskeletal disorders that affect the joints. Despite high prevalence rates, etiological factors involved in these disorders remain largely unknown. Dissecting the molecular aspects of these disorders will significantly contribute to improving their diagnosis and clinical management. In order to identify proteins that are differentially expressed between these two conditions, a quantitative proteomic profiling of synovial fluid obtained from rheumatoid arthritis and osteoarthritis patients was carried out by using iTRAQ labeling followed by high resolution mass spectrometry analysis.

Results

We have identified 575 proteins out of which 135 proteins were found to be differentially expressed by ≥3-fold in the synovial fluid of rheumatoid arthritis and osteoarthritis patients. Proteins not previously reported to be associated with rheumatoid arthritis including, coronin-1A (CORO1A), fibrinogen like-2 (FGL2), and macrophage capping protein (CAPG) were found to be upregulated in rheumatoid arthritis. Proteins such as CD5 molecule-like protein (CD5L), soluble scavenger receptor cysteine-rich domain-containing protein (SSC5D), and TTK protein kinase (TTK) were found to be upregulated in the synovial fluid of osteoarthritis patients. We confirmed the upregulation of CAPG in rheumatoid arthritis synovial fluid by multiple reaction monitoring assay as well as by Western blot. Pathway analysis of differentially expressed proteins revealed a significant enrichment of genes involved in glycolytic pathway in rheumatoid arthritis.

Conclusions

We report here the largest identification of proteins from the synovial fluid of rheumatoid arthritis and osteoarthritis patients using a quantitative proteomics approach. The novel proteins identified from our study needs to be explored further for their role in the disease pathogenesis of rheumatoid arthritis and osteoarthritis.

Sartaj Ahmad and Raja Sekhar Nirujogi contributed equally to this article.

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Most cited references 77

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Gene Ontology: tool for the unification of biology

Michael Ashburner, Catherine A. Ball, Judith Blake … (2002)

Genomic sequencing has made it clear that a large fraction of the genes specifying the core biological functions are shared by all eukaryotes. Knowledge of the biological role of such shared proteins in one organism can often be transferred to other organisms. The goal of the Gene Ontology Consortium is to produce a dynamic, controlled vocabulary that can be applied to all eukaryotes even as knowledge of gene and protein roles in cells is accumulating and changing. To this end, three independent ontologies accessible on the World-Wide Web (http://www.geneontology.org) are being constructed: biological process, molecular function and cellular component.

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The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis.

D McShane, D. Bloch, Emma Healey … (1988)

The revised criteria for the classification of rheumatoid arthritis (RA) were formulated from a computerized analysis of 262 contemporary, consecutively studied patients with RA and 262 control subjects with rheumatic diseases other than RA (non-RA). The new criteria are as follows: 1) morning stiffness in and around joints lasting at least 1 hour before maximal improvement; 2) soft tissue swelling (arthritis) of 3 or more joint areas observed by a physician; 3) swelling (arthritis) of the proximal interphalangeal, metacarpophalangeal, or wrist joints; 4) symmetric swelling (arthritis); 5) rheumatoid nodules; 6) the presence of rheumatoid factor; and 7) radiographic erosions and/or periarticular osteopenia in hand and/or wrist joints. Criteria 1 through 4 must have been present for at least 6 weeks. Rheumatoid arthritis is defined by the presence of 4 or more criteria, and no further qualifications (classic, definite, or probable) or list of exclusions are required. In addition, a "classification tree" schema is presented which performs equally as well as the traditional (4 of 7) format. The new criteria demonstrated 91-94% sensitivity and 89% specificity for RA when compared with non-RA rheumatic disease control subjects.

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Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis.

Kristine A. Kuhn, William P Arend, V Michael Holers … (2006)

Antibodies against citrullinated proteins are specific and predictive markers for rheumatoid arthritis although the pathologic relevance of these antibodies remains unclear. To investigate the significance of these autoantibodies, collagen-induced arthritis (CIA) in mice was used to establish an animal model of antibody reactivity to citrullinated proteins. DBA/1J mice were immunized with bovine type II collagen (CII) at days 0 and 21, and serum was collected every 7 days for analysis. Antibodies against both CII and cyclic citrullinated peptide, one such citrullinated antigen, appeared early after immunization, before joint swelling was observed. Further, these antibodies demonstrated specific binding to citrullinated filaggrin in rat esophagus by indirect immunofluorescence and citrullinated fibrinogen by Western blot. To evaluate the role of immune responses to citrullinated proteins in CIA, mice were tolerized with a citrulline-containing peptide, followed by antigen challenge with CII. Tolerized mice demonstrated significantly reduced disease severity and incidence compared with controls. We also identified novel murine monoclonal antibodies specific to citrullinated fibrinogen that enhanced arthritis when coadministered with a submaximal dose of anti-CII antibodies and bound targets within the inflamed synovium of mice with CIA. These results demonstrate that antibodies against citrullinated proteins are centrally involved in the pathogenesis of autoimmune arthritis.

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Author and article information

Journal

Journal ID (nlm-ta): Clin Proteomics

Journal ID (iso-abbrev): Clin Proteomics

Title: Clinical proteomics

Publisher: Springer

ISSN (Print): 1542-6416

ISSN (Electronic): 1559-0275

Publication date Collection: 2014

Publication date (Electronic): 6 January 2014

Volume: 11

Issue: 1

Page: 1

Affiliations

[1 ]Institute of Bioinformatics, International Technology Park, Bangalore 560066, India

[2 ]Department of Biotechnology, Kuvempu University, Shankaraghatta 577451, India

[3 ]Amrita School of Biotechnology, Amrita Vishwa Vidyapeetham, Kollam 690525, India

[4 ]Manipal University, Madhava Nagar, Manipal 576104, India

[5 ]Centre for Excellence in Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry 605014, India

[6 ]Rajiv Gandhi University of Health Sciences, Bangalore 560041, India

[7 ]Department of Internal Medicine, Armed Forces Medical College, Pune 411040, India

[8 ]Department of Rheumatology, Fortis Hospitals, Bangalore 560076, India

[9 ]Department of Rheumatology, Command Airforce Hospital, Bangalore 560008, India

[10 ]Laboratory for Integrated Bioinformatics, RIKEN Center for Integrative Medical Sciences (IMS-RCAI), Yokohama Institute, Kanagawa 230-0045, Japan

[11 ]McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

[12 ]Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

[13 ]Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

[14 ]Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

Article

Publisher ID: 1559-0275-11-1

DOI: 10.1186/1559-0275-11-1

PMC ID: 3918105

PubMed ID: 24393543

SO-VID: f29e7bd5-7583-4208-ad46-daca84eed47e

License:

This is an open access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Differential proteomic analysis of synovial fluid from rheumatoid arthritis and osteoarthritis patients

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Abstract

Background

Results

Conclusions

Related collections

Core Readings in Statistical Mediation Analysis

Most cited references 77

Gene Ontology: tool for the unification of biology

The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis.

Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis.

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