Urine Proteomics Profiling and Functional Characterization of Knee Osteoarthritis Using iTRAQ Technology

Abstract

Osteoarthritis (OA) is a degenerative chronic disease affecting the whole joint structures. With the increment in life expectancy and aging population, OA has become one of the largest socioeconomic burdens, associated with pain and loss of joint function. However, early laboratory tests of OA are still lacking. Therefore, new diagnostic tests for this disease are urgently needed. In this study, to gain an insight into the pathogenesis and the potential biomarkers of OA, we implemented a comparative urine proteomics study on OA patients and health people using iTRAQ-based mass spectrometry technology. Western blotting was used to validate the relative changes in urine protein levels for four of the identified proteins. We constructed a comprehensive urine proteome profile of the OA patients and identified 102 proteins differently changed in abundance. Forty-six proteins were upregulated and 56 proteins were significantly downregulated in OA patients. Furthermore, the proteins, COL-4, MMP9, adiponectin, and BBOX1 were validated through Western blots, which can serve as valuable candidate biomarkers and help to illustrate the pathogenesis of OA. These findings may provide clues for promising biomarkers for the early diagnosis and also offer a theoretical basis for the early treatment of OA.

Key words

osteoarthritis - urine proteomics - iTRAQ, biomarker

• References

• 1 Tang X, Wang S, Zhan S. et al. The prevalence of symptomatic knee osteoarthritis in China: Results from the China Health and Retirement Longitudinal Study. Arthritis Rheumatol 2016; 68: 648-653
• 2 Helmick CG, Felson DT, Lawrence RC. et al. Estimates of the prevalence of arthritis and other rheumatic conditions in the United States. Part I. Arthritis Rheum 2008; 58: 15-25
• 3 Lawrence RC, Felson DT, Helmick CG. et al. Estimates of the prevalence of arthritis and other rheumatic conditions in the United States. Part II. Arthritis Rheum 2008; 58: 26-35
• 4 Hiligsmann M, Cooper C, Arden N. et al. Health economics in the field of osteoarthritis: an expert's consensus paper from the European Society for Clinical and Economic Aspects of Osteoporosis and Osteoarthritis (ESCEO). Semin Arthritis Rheum 2013; 43: 303-313
• 5 Bournia VK, Kitas G, Protogerou AD. et al. Impact of non-steroidal anti-inflammatory drugs on cardiovascular risk: Is it the same in osteoarthritis and rheumatoid arthritis?. Mod Rheumatol 2017; 27: 559-569
• 6 Martel-Pelletier J, Barr AJ, Cicuttini FM. et al. Osteoarthritis. Nat Rev Dis Primers 2016; 2: 16072
• 7 Yu SP, Hunter DJ.. Managing osteoarthritis. Aust Prescr 2015; 38: 115-119
• 8 Munoz J, Heck AJ. From the human genome to the human proteome. Angew Chem Int Ed 2014; 53: 10864-10866
• 9 Mobasheri A, Bay-Jensen AC, van Spil WE. et al. Osteoarthritis Year in Review 2016: Biomarkers (biochemical markers). Osteoarthritis Cartilage 2017; 25: 199-208
• 10 Liao W, Li Z, Li T. et al. Proteomic analysis of synovial fluid in osteoarthritis using SWATHmass spectrometry. Mol Med Rep 2018; 17: 2827-2836
• 11 Beasley-Green A.. Urine Proteomics in the Era of Mass Spectrometry. Int Neurourol J 2016; 20: S70-S75
• 12 Nunez EV, Domont GB, Nogueira FC.. iTRAQ-Based shotgun proteomics approach for relative protein quantification. Methods Mol Biol 2017; 1546: 267-274
• 13 Nemirovskiy O, Li WW, Szekely-Klepser G.. Design and validation of an immunoaffinity LC-MS/MS assay for the quantification of a collagen type II neoepitope peptide in human urine: application as a biomarker of osteoarthritis. Methods Mol Biol 2010; 641: 253-270
• 14 Xiaoyu L, Weiyuan Z, Ping J. et al. Comparative serum proteomic analysis of adenomyosis using the isobaric tags for relative and absolute quantitation technique. Fertil Steril 2013; 100: 505-510
• 15 Saxby DJ, Llyod DG. Osteoarthritis and cartilage. Osteoarthritis year in review 2016: Mechanics. Osteoarthritis Cartilage 2017; 25: 190-198
• 16 Zeng GQ, Chen AB, Li W. et al. High MMP-1, MMP-2, and MMP-9 protein levels in osteoarthritis. Genet Mol Res 2015; 14: 14811-14822
• 17 Gong Z, Tas E, Yakar S. et al. Hepatic lipid metabolism and non-alcoholic fatty liver disease in aging. Mol Cell Endocrinol 2017; 455: 115-130
• 18 Cuzdan Coskun N, Ay S, Evcik FD. et al. Adiponectin: Is it a biomarker for assessing the disease severity in knee osteoarthritis patients?. Int J Rheum Dis 2017; 20: 1942-1949
• 19 Arita Y, Kihara S, Ouchi N. et al. Paradoxical decrease of an adipose-specific protein, adiponectin, in obesity. Biochem Biophys Res Commun 1999; 257: 79-83
• 20 Thijssen E, van Caam A, van der Kraan PM.. Obesity and osteoarthritis, more than just wear and tear: pivotal roles for inflamed adipose tissue and dyslipidaemia in obesity-induced osteoarthritis. Rheumatology (Oxford) 2015; 54: 588-600
• 21 Goto N, Okazaki K, Akasaki Y. et al. Single intra-articular injection of fluvastatin-PLGA microspheres reduces cartilage degradation in rabbits with experimental osteoarthritis. J Orthop Res 2017; 35: 2465-2475