Skip to main content

Advertisement

SpringerLink
Log in

Identification of Potential Serum Biomarkers for Rheumatoid Arthritis by High-Resolution Quantitative Proteomic Analysis

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

The aim of this study was to find serum biomarkers of rheumatoid arthritis (RA) by high-resolution proteomic analysis. Low-abundance proteins from pooling serum sample of early RA patients and healthy controls were enriched using ProteoMiner™ enrichment kits. The enriched proteins were separated on SDS-PAGE, digested by trypsin, labeled with tandem mass tag (TMT) reagents, and desalted by C18 stage tip column. Then, the labeled peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with nano-LC combined with Orbitrap Q Exactive mass spectrometer, and experiments were carried out three times using different specimens, and differentially expressed proteins were screened by intensity ratios of identified peptides. Enzyme-linked immunosorbent assays (ELISAs) were performed to confirm differentially expressed proteins. Twenty-six proteins were found differentially expressed in RA serum by high-resolution proteomic analysis. Among them, levels of thrombospondin-1, ficolin-2, isoform 10 of fibronectin, and apolipoprotein E were higher in RA patients than in healthy controls (RA/healthy control (HC) ≥ 1.5, p < 0.05), while levels of angiotensinogen, paraoxonase/arylesterase 1, isoform E of proteoglycan 4, and plasminogen were significantly lower (RA/HC ≤ 0.67, p < 0.05). Further study by ELISA showed a higher level of ficolin-2 in the serum of RA patients compared to healthy controls; the level of ficolin-2 was found to be positively correlated with swollen joint counts (SJCs), disease activity score (DAS28), rheumatoid factor, and IgM in RA patients and with DAS28 and IgM in early RA patients through statistical analysis. The results of this study suggest that ficolin-2, as a newly screened biomarker by high-resolution quantitative proteomic analysis, offers the potentiality to become a diagnostic or disease evaluation tool in RA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

RA:

Rheumatoid arthritis

ELISA:

Enzyme-linked immunosorbent assay

SJCs:

Swollen joint counts

TJCs:

Tender joint counts

DAS:

Disease activity score

MALDI-TOF:

Matrix-assisted laser desorption/ionization time of flight

TMT:

Tandem mass tag

SSc:

Systemic sclerosis

OA:

Osteoarthritis

BD:

Behcet’s disease

HC:

Healthy control

CRP:

C-reactive protein

ESR:

Erythrocyte sedimentation rate

2-DE:

Two-dimensional electrophoresis

TSP-1:

Thrombospondin-1

FCN-2:

Ficolin-2

REFERENCES

  1. Li, R., J. Sun, L.M. Ren, H.Y. Wang, W.H. Liu, X.W. Zhang, S. Chen, R. Mu, J. He, Y. Zhao, L. Long, Y.Y. Liu, X. Liu, X.L. Lu, Y.H. Li, S.Y. Wang, S.S. Pan, C. Li, and Z.G. Li. 2012. Epidemiology of eight common rheumatic diseases in China: a large-scale cross-sectional survey in Beijing. Rheumatology (Oxford) 51: 721–729.

    Article  Google Scholar 

  2. Panayi, G.S. 2005. B cells: a fundamental role in the pathogenesis of rheumatoid arthritis? Rheumatology (Oxford) 44(Suppl 2): ii3–ii7.

    CAS  Google Scholar 

  3. van Gaalen, F.A., S.P. Linn-Rasker, W.J. van Venrooij, B.A. de Jong, F.C. Breedveld, C.L. Verweij, R.E. Toes, and T.W. Huizinga. 2004. Autoantibodies to cyclic citrullinated peptides predict progression to rheumatoid arthritis in patients with undifferentiated arthritis: a prospective cohort study. Arthritis and Rheumatism 50: 709–715.

    Article  PubMed  Google Scholar 

  4. Canhao, H., A.M. Rodrigues, A.F. Mourao, F. Martins, M.J. Santos, J. Canas-Silva, J. Polido-Pereira, J.A. Pereira Silva, J.A. Costa, D. Araujo, C. Silva, H. Santos, C. Duarte, J.A. da Silva, F.M. Pimentel-Santos, J.C. Branco, E.W. Karlson, J.E. Fonseca, and D.H. Solomon. 2012. Comparative effectiveness and predictors of response to tumour necrosis factor inhibitor therapies in rheumatoid arthritis. Rheumatology (Oxford) 51: 2020–2026.

    Article  CAS  Google Scholar 

  5. Bykerk, V.P., S. Jamal, G. Boire, C.A. Hitchon, B. Haraoui, J.E. Pope, J.C. Thorne, Y. Sun, and E.C. Keystone. 2012. The Canadian Early Arthritis Cohort (CATCH): patients with new-onset synovitis meeting the 2010 ACR/EULAR classification criteria but not the 1987 ACR classification criteria present with less severe disease activity. Journal of Rheumatology 39: 2071–2080.

    Article  PubMed  Google Scholar 

  6. Jung, Y.O., and H.A. Kim. 2012. Recent paradigm shifts in the diagnosis and treatment of rheumatoid arthritis. Korean Journal of Internal Medicine 27: 378–387.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  7. Apweiler, R., C. Aslanidis, T. Deufel, A. Gerstner, J. Hansen, D. Hochstrasser, R. Kellner, M. Kubicek, F. Lottspeich, E. Maser, H.W. Mewes, H.E. Meyer, S. Mullner, W. Mutter, M. Neumaier, P. Nollau, H.G. Nothwang, F. Ponten, A. Radbruch, K. Reinert, G. Rothe, H. Stockinger, A. Tarnok, M.J. Taussig, A. Thiel, J. Thiery, M. Ueffing, G. Valet, J. Vandekerckhove, W. Verhuven, C. Wagener, O. Wagner, and G. Schmitz. 2009. Approaching clinical proteomics: current state and future fields of application in fluid proteomics. Clinical Chemistry and Laboratory Medicine 47: 724–744.

    Article  PubMed  CAS  Google Scholar 

  8. Wang, J.G., W.D. Xu, W.T. Zhai, Y. Li, J.W. Hu, B. Hu, M. Li, L. Zhang, W. Guo, J.P. Zhang, L.H. Wang, and B.H. Jiao. 2012. Disorders in angiogenesis and redox pathways are main factors contributing to the progression of rheumatoid arthritis: a comparative proteomics study. Arthritis and Rheumatism 64: 993–1004.

    Article  PubMed  CAS  Google Scholar 

  9. Liao, H., J. Wu, E. Kuhn, W. Chin, B. Chang, M.D. Jones, S. O’Neil, K.R. Clauser, J. Karl, F. Hasler, R. Roubenoff, W. Zolg, and B.C. Guild. 2004. Use of mass spectrometry to identify protein biomarkers of disease severity in the synovial fluid and serum of patients with rheumatoid arthritis. Arthritis and Rheumatism 50: 3792–3803.

    Article  PubMed  CAS  Google Scholar 

  10. Jin, E.H., S.C. Shim, H.G. Kim, S.C. Chae, and H.T. Chung. 2009. Polymorphisms of COTL1 gene identified by proteomic approach and their association with autoimmune disorders. Experimental and Molecular Medicine 41: 354–361.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  11. Makarov, A., E. Denisov, O. Lange, and S. Horning. 2006. Dynamic range of mass accuracy in LTQ Orbitrap hybrid mass spectrometer. Journal of The American Society for Mass Spectrometry 17: 977–982.

    Article  PubMed  CAS  Google Scholar 

  12. Makarov, A., E. Denisov, A. Kholomeev, W. Balschun, O. Lange, K. Strupat, and S. Horning. 2006. Performance evaluation of a hybrid linear ion trap/orbitrap mass spectrometer. Analytical Chemistry 78: 2113–2120.

    Article  PubMed  CAS  Google Scholar 

  13. Raso, C., C. Cosentino, M. Gaspari, N. Malara, X. Han, D. McClatchy, S.K. Park, M. Renne, N. Vadala, U. Prati, G. Cuda, V. Mollace, F. Amato, and J.R. Yates 3rd. 2012. Characterization of breast cancer interstitial fluids by TmT labeling, LTQ-Orbitrap Velos mass spectrometry, and pathway analysis. Journal of Proteome Research 11: 3199–3210.

    Article  PubMed  CAS  Google Scholar 

  14. Aletaha, D., T. Neogi, A.J. Silman, J. Funovits, D.T. Felson, C.O. Bingham 3rd, N.S. Birnbaum, G.R. Burmester, V.P. Bykerk, M.D. Cohen, B. Combe, K.H. Costenbader, M. Dougados, P. Emery, G. Ferraccioli, J.M. Hazes, K. Hobbs, T.W. Huizinga, A. Kavanaugh, J. Kay, T.K. Kvien, T. Laing, P. Mease, H.A. Menard, L.W. Moreland, R.L. Naden, T. Pincus, J.S. Smolen, E. Stanislawska-Biernat, D. Symmons, P.P. Tak, K.S. Upchurch, J. Vencovsky, F. Wolfe, and G. Hawker. 2010. 2010 rheumatoid arthritis classification criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Annals of the Rheumatic Diseases 69: 1580–1588.

    Article  PubMed  Google Scholar 

  15. (1980). Preliminary criteria for the classification of systemic sclerosis (scleroderma). Subcommittee for scleroderma criteria of the American Rheumatism Association Diagnostic and Therapeutic Criteria Committee. Arthritis Rheum 23:581-590.

  16. International Study Group for Behcet’s Disease. 1990. Criteria for diagnosis of Behcet’s disease. Lancet 335: 1078–1080.

    Google Scholar 

  17. Altman, R.D. 1995. The classification of osteoarthritis. Journal of Rheumatology. Supplement 43: 42–43.

    CAS  Google Scholar 

  18. Hu, S., J.A. Loo, and D.T. Wong. 2007. Human saliva proteome analysis. Annals of the New York Academy of Sciences 1098: 323–329.

    Article  PubMed  CAS  Google Scholar 

  19. Hu, S., J.A. Loo, and D.T. Wong. 2006. Human body fluid proteome analysis. Proteomics 6: 6326–6353.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  20. Serada, S., M. Fujimoto, A. Ogata, F. Terabe, T. Hirano, H. Iijima, S. Shinzaki, T. Nishikawa, T. Ohkawara, K. Iwahori, N. Ohguro, T. Kishimoto, and T. Naka. 2010. iTRAQ-based proteomic identification of leucine-rich alpha-2 glycoprotein as a novel inflammatory biomarker in autoimmune diseases. Annals of the Rheumatic Diseases 69: 770–774.

    Article  PubMed  CAS  Google Scholar 

  21. Zheng, X., S.L. Wu, M. Hincapie, and W.S. Hancock. 2009. Study of the human plasma proteome of rheumatoid arthritis. Journal of Chromatography A 1216: 3538–3545.

    Article  PubMed  CAS  Google Scholar 

  22. Falgarone, G., A. Essabbani, F. Dumont, N. Cagnard, S. Mistou, and G. Chiocchia. 2011. Implication of clusterin in TNF-alpha response of rheumatoid synovitis: lesson from in vitro knock-down of clusterin in human synovial fibroblast cells. Physiological Genomics 44: 229–235.

    Article  PubMed  Google Scholar 

  23. Przybysz, M., K. Borysewicz, and I. Katnik-Prastowska. 2013. Fibronectin molecular status determination useful to differentiate between rheumatoid arthritis and systemic lupus erythematosus patients. Rheumatology International 33: 37–43.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  24. Biro, J., Z. Balogh, M. Szongoth, A. Megyeri, M. Bely, and A. Falus. 1992. Molecular genetic, laboratory and clinical study of serum amyloid P. Orvosi Hetilap 133: 841–845.

    PubMed  CAS  Google Scholar 

  25. Friese, M.A., T. Manuelian, S. Junnikkala, J. Hellwage, S. Meri, H.H. Peter, D.L. Gordon, H. Eibel, and P.F. Zipfel. 2003. Release of endogenous anti-inflammatory complement regulators FHL-1 and factor H protects synovial fibroblasts during rheumatoid arthritis. Clinical and Experimental Immunology 132: 485–495.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  26. Jacobson, G.A., S.J. Ives, C. Narkowicz, and G. Jones. 2012. Plasma glutathione peroxidase (GSH-Px) concentration is elevated in rheumatoid arthritis: a case-control study. Clinical Rheumatology 31: 1543–1547.

    Article  PubMed  Google Scholar 

  27. van Beers, J.J., C.M. Schwarte, J. Stammen-Vogelzangs, E. Oosterink, B. Bozic, and G.J. Pruijn. 2013. The rheumatoid arthritis synovial fluid citrullinome reveals novel citrullinated epitopes in apolipoprotein E, myeloid nuclear differentiation antigen, and beta-actin. Arthritis and Rheumatism 65: 69–80.

    Article  PubMed  Google Scholar 

  28. Ichijo, H., L. Ronnstrand, K. Miyagawa, H. Ohashi, C.H. Heldin, and K. Miyazono. 1991. Purification of transforming growth factor-beta 1 binding proteins from porcine uterus membranes. Journal of Biological Chemistry 266: 22459–22464.

    PubMed  CAS  Google Scholar 

  29. Endo, Y., M. Matsushita, and T. Fujita. 2011. The role of ficolins in the lectin pathway of innate immunity. International Journal of Biochemistry and Cell Biology 43: 705–712.

    Article  PubMed  CAS  Google Scholar 

  30. Endo, Y., N. Nakazawa, D. Iwaki, M. Takahashi, M. Matsushita, and T. Fujita. 2010. Interactions of ficolin and mannose-binding lectin with fibrinogen/fibrin augment the lectin complement pathway. Journal of Innate Immunity 2: 33–42.

    Article  PubMed  CAS  Google Scholar 

  31. Matsushita, M. 2010. Ficolins: complement-activating lectins involved in innate immunity. Journal of Innate Immunity 2: 24–32.

    Article  PubMed  CAS  Google Scholar 

  32. Krarup, A., U.B. Sorensen, M. Matsushita, J.C. Jensenius, and S. Thiel. 2005. Effect of capsulation of opportunistic pathogenic bacteria on binding of the pattern recognition molecules mannan-binding lectin, L-ficolin, and H-ficolin. Infection and Immunity 73: 1052–1060.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  33. Aoyagi, Y., E.E. Adderson, C.E. Rubens, J.F. Bohnsack, J.G. Min, M. Matsushita, T. Fujita, Y. Okuwaki, and S. Takahashi. 2008. L-Ficolin/mannose-binding lectin-associated serine protease complexes bind to group B streptococci primarily through N-acetylneuraminic acid of capsular polysaccharide and activate the complement pathway. Infection and Immunity 76: 179–188.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  34. Kuraya, M., Z. Ming, X. Liu, M. Matsushita, and T. Fujita. 2005. Specific binding of L-ficolin and H-ficolin to apoptotic cells leads to complement activation. Immunobiology 209: 689–697.

    Article  PubMed  CAS  Google Scholar 

  35. Zhang, J., J. Koh, J. Lu, S. Thiel, B.S. Leong, S. Sethi, C.Y. He, B. Ho, and J.L. Ding. 2009. Local inflammation induces complement crosstalk which amplifies the antimicrobial response. PLoS Pathogens 5: e1000282.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Nakaya, H.I., J. Gardner, Y.S. Poo, L. Major, B. Pulendran, and A. Suhrbier. 2012. Gene profiling of Chikungunya virus arthritis in a mouse model reveals significant overlap with rheumatoid arthritis. Arthritis and Rheumatism 64: 3553–3563.

    Article  PubMed  CAS  Google Scholar 

  37. Pender, M.P. 2012. CD8+ T-cell deficiency, Epstein-Barr virus infection, vitamin D deficiency, and steps to autoimmunity: a unifying hypothesis. Autoimmune Diseases 2012: 189096.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Ammitzboll, C.G., S. Thiel, T. Ellingsen, B. Deleuran, A. Jorgensen, J.C. Jensenius, and K. Stengaard-Pedersen. 2012. Levels of lectin pathway proteins in plasma and synovial fluid of rheumatoid arthritis and osteoarthritis. Rheumatology International 32: 1457–1463.

    Article  PubMed  CAS  Google Scholar 

  39. Vander Cruyssen, B., L. Nuytinck, L. Boullart, D. Elewaut, W. Waegeman, M. Van Thielen, E. De Meester, K. Lebeer, R. Rossau, and F. De Keyser. 2007. Polymorphisms in the ficolin 1 gene (FCN1) are associated with susceptibility to the development of rheumatoid arthritis. Rheumatology (Oxford) 46: 1792–1795.

    Article  CAS  Google Scholar 

  40. Kolarov, Z., R. Stoilov, M. Baleva, K. Nikolov, I. Sheitanov, M. Iancheva, N. Tsacheva, and S. Radenska-Lopovok. 2000. Alpha-2 macroglobulin in serum and synovial fluid of patients with rheumatoid arthritis and osteoarthrosis. Terapevticheskiĭ Arkhiv 72: 17–19.

    PubMed  CAS  Google Scholar 

  41. Isik, A., S.S. Koca, B. Ustundag, H. Celik, and A. Yildirim. 2007. Paraoxonase and arylesterase levels in rheumatoid arthritis. Clinical Rheumatology 26: 342–348.

    Article  PubMed  CAS  Google Scholar 

  42. Li, J., Y. Guo, R. Holmdahl, and T. Ny. 2005. Contrasting roles of plasminogen deficiency in different rheumatoid arthritis models. Arthritis and Rheumatism 52: 2541–2548.

    Article  PubMed  CAS  Google Scholar 

  43. Busso, N., V. Peclat, K. Van Ness, E. Kolodziesczyk, J. Degen, T. Bugge, and A. So. 1998. Exacerbation of antigen-induced arthritis in urokinase-deficient mice. Journal of Clinical Investigation 102: 41–50.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  44. Metsarinne, K.P., D.C. Nordstrom, Y.T. Konttinen, A.M. Teppo, and F.Y. Fyhrquist. 1992. Plasma interleukin-6 and renin substrate in reactive arthritis, rheumatoid arthritis, and systemic lupus erythematosus. Rheumatology International 12: 93–96.

    Article  PubMed  CAS  Google Scholar 

Download references

ACKNOWLEDGMENTS

The research was supported by Grant 81030057 from National Natural Science Foundation, the National Basic Research Program of China(973 program 2010CB529100), and Grant 31000409 from National Natural Science Foundation.

Author information

Authors and Affiliations

  1. Department of Rheumatology and Immunology, People’s Hospital, Peking University, 11 Xizhimen South St., 100044, Beijing, China

    Xiaolin Sun, Yuhui Li & Zhanguo Li

  2. Department of Rheumatology and Immunology, Beijing Hospital, 1 Dahua Road, 100730, Beijing, China

    Yongjing Cheng & Cibo Huang

  3. School of Life Sciences, Tsinghua University, 100084, Beijing, China

    Yuling Chen & Haiteng Deng

Authors
  1. Yongjing Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuling Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaolin Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuhui Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cibo Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Haiteng Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zhanguo Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Haiteng Deng or Zhanguo Li.

Additional information

Yongjing Cheng, Yuling Chen and Xiaolin Sun contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cheng, Y., Chen, Y., Sun, X. et al. Identification of Potential Serum Biomarkers for Rheumatoid Arthritis by High-Resolution Quantitative Proteomic Analysis. Inflammation 37, 1459–1467 (2014). https://doi.org/10.1007/s10753-014-9871-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-014-9871-8

KEY WORDS

Search

Navigation