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Home / Equine Research Bank / Equine Vet J / Metabolism and global protein glycosylation are differential...
Equine veterinary journal2021; 54(2); 323-333; doi: 10.1111/evj.13440

Metabolism and global protein glycosylation are differentially expressed in healthy and osteoarthritic equine carpal synovial fluid.

Abstract: Carpal osteochondral fragmentation and subsequent post-traumatic osteoarthritis (PTOA) are leading causes of wastage in the equine athlete. Identification of synovial fluid biomarkers could contribute to the diagnosis and understanding of osteoarthritis (OA) pathophysiology. Objective: The aim of this study was to identify differentially expressed metabolic and glycosylation pathways in synovial fluid from healthy horses and horses with naturally occurring carpal OA. Methods: Cross-sectional, in vivo metabolomics and glycomics study. Methods: In cohort 1, carpal synovial fluid (n = 12 horses; n = 6 healthy, n = 6 OA) was analysed using high-resolution liquid chromatography mass spectrometry (LC-MS). In cohort 2 (n = 40 horses; n = 20 healthy, n = 20 OA), carpal synovial fluid was analysed using lectin microarrays and a lubricin sandwich ELISA. Results: Metabolomic analysis identified >4900 LC-MS features of which 84 identifiable metabolites were differentially expressed (P < .05) between healthy and OA joints, including key pathways related to inflammation (histidine and tryptophan metabolism), oxidative stress (arginine biosynthesis) and collagen metabolism (lysine metabolism). Principle Component Analysis and Partial Least Squares Discriminant Analysis demonstrated separation between healthy and OA synovial fluid. Lectin microarrays identified distinct glycosylation patterns between healthy and OA synovial fluid, including increased Core 1/Core 3 O-glycosylation, increased α-2,3 sialylation and decreased α-1,2 fucosylation in OA. O-glycans predominated over N-glycans in all synovial fluid samples, and synovial fluid lubricin was increased in OA joints as compared to controls. Conclusions: The sample size in cohort 1 was limited, and there is inherent variation in severity and duration of joint injury in naturally occurring OA. However, LC-MS identified up to 5000 unique features. Conclusions: These data suggest new potential diagnostic and therapeutic targets for equine OA. Future targeted metabolomic and glycomic studies should be performed to verify these results. Lectin microarrays could be investigated as a potential screening tool for the diagnosis and therapeutic monitoring of equine OA.
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Publication Date: 2021-03-18 PubMed ID: 33587757PubMed Central: PMC8364562DOI: 10.1111/evj.13440Google Scholar: Lookup
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Summary

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The study explores the differences in metabolic and glycosylation pathways in the synovial fluid of healthy horses compared to horses with osteoarthritis focusing on equine athletes. The aim was to uncover potential biomarkers towards improving diagnosis and understanding the disease’s pathophysiology.

Objectives and Methods of the Study

  • The main objective of this study was to identify any differences in metabolism and protein glycosylation in synovial fluid from healthy horses versus those with osteoarthritis (OA).
  • Two cohorts of horses were used for the study. In the first group, 12 horses’ synovial fluid, six healthy and six with OA, was analyzed using high-resolution liquid chromatography mass spectrometry (LC-MS).
  • For the second group, the synovial fluid from 40 horses, 20 healthy and 20 with OA, was examined using lectin microarrays and lubricin sandwich ELISA.

Findings of the Study

  • Metabolomic analysis discovered more than 4900 features, with 84 identifiable and differentially expressed metabolites (changes in their levels) between healthy and OA synovial fluid.
  • This included noticeable changes linked to inflammation (histidine and tryptophan metabolism), oxidative stress (arginine biosynthesis), and collagen metabolism (lysine metabolism).
  • The statistical tests, Principle Component Analysis and Partial Least Squares Discriminant Analysis, showed a clear distinction between healthy and OA synovial fluid samples.
  • The lectin microarray test identified that different protein glycosylation patterns exist between healthy and OA synovial fluid.
  • In particular, in OA synovial fluid, there was an increase in Core 1/Core 3 O-glycosylation and α-2,3 sialylation along with a decrease in α-1,2 fucosylation.
  • O-glycans predominated over N-glycans in all equine synovial fluid samples, and the lubricin levels were found to be increased in the OA joints compared to the healthy ones.

Conclusions and Recommendations

  • The study faced some limitations. The number of equine samples in the first group was limited, and there were also variable severity and duration of joint damage in the case of naturally occurring osteoarthritis.
  • Despite these limitations, new potential diagnostic and therapeutic targets for treating equine OA have been identified.
  • To validate these findings, the authors recommend targeted metabolomic and glycomic studies in future research.
  • The lectin microarray method also holds promise as a potential screening tool to facilitate early diagnosis and therapeutic monitoring of osteoarthritis in equine athletes.

Cite This Article

APA
Noordwijk KJ, Qin R, Diaz-Rubio ME, Zhang S, Su J, Mahal LK, Reesink HL. (2021). Metabolism and global protein glycosylation are differentially expressed in healthy and osteoarthritic equine carpal synovial fluid. Equine Vet J, 54(2), 323-333. https://doi.org/10.1111/evj.13440

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 54
Issue: 2
Pages: 323-333

Researcher Affiliations

Noordwijk, Kira J
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Qin, Rui
  • Department of Chemistry, New York University, New York, NY, USA.
  • Department of Chemistry, University of Alberta, Edmonton, AB, Canada.
Diaz-Rubio, Maria E
  • Proteomics and Metabolomics Facility, Institute of Biotechnology, Cornell University, Ithaca, NY, USA.
Zhang, Sheng
  • Proteomics and Metabolomics Facility, Institute of Biotechnology, Cornell University, Ithaca, NY, USA.
Su, Jin
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Mahal, Lara K
  • Department of Chemistry, New York University, New York, NY, USA.
  • Department of Chemistry, University of Alberta, Edmonton, AB, Canada.
Reesink, Heidi L
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.

MeSH Terms

  • Animals
  • Biomarkers / metabolism
  • Cross-Sectional Studies
  • Glycosylation
  • Horse Diseases / metabolism
  • Horses
  • Joint Diseases / veterinary
  • Osteoarthritis / metabolism
  • Osteoarthritis / veterinary
  • Synovial Fluid / metabolism

Grant Funding

  • R24 GM08291 / Foundation for the National Institutes of Health
  • R24 GM082910 / NIGMS NIH HHS
  • T32 GM008291 / NIGMS NIH HHS
  • R24 GM08291 / Cornell Veterinary Biobank
  • K08 AR068469 / NIAMS NIH HHS
  • K08AR068469 / NIAMS NIH HHS

Conflict of Interest Statement

CONFLICT OF INTERESTS. No competing interests have been declared.

References

This article includes 44 references
  1. McIlwraith CW. Arthroscopic surgery for osteochondral chip fragments and other lesions not requiring internal fixation in the carpal and fetlock joints of the equine athlete: what have we learned in 20 years?. Clin Tech Equine Pract 2002;1(4):200–10.
  2. McIlwraith CW, Kawcak CE, Frisbie DD, Little CB, Clegg PD, Peffers MJ, Karsdal MA, Ekman S, Laverty S, Slayden RA, Sandell LJ, Lohmander LS, Kraus VB. Biomarkers for equine joint injury and osteoarthritis.. J Orthop Res 2018 Mar;36(3):823-831.
    pubmed: 28921609doi: 10.1002/jor.23738google scholar: lookup
  3. McIlwraith CW, Clegg PD. Science in brief: Report on the Havemeyer Foundation workshop on equine musculoskeletal biomarkers--current knowledge and future needs.. Equine Vet J 2014 Nov;46(6):651-3.
    pubmed: 25319159doi: 10.1111/evj.12339google scholar: lookup
  4. Skiöldebrand E, Ekman S, Mattsson Hultén L, Svala E, Björkman K, Lindahl A, Lundqvist A, Önnerfjord P, Sihlbom C, Rüetschi U. Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis.. Equine Vet J 2017 Sep;49(5):662-667.
    pmc: PMC5573946pubmed: 28097685doi: 10.1111/evj.12666google scholar: lookup
  5. Anderson JR, Smagul A, Simpson D, Clegg PD, Rubio-Martinez LM, Peffers MJ. The synovial fluid proteome differentiates between septic and nonseptic articular pathologies.. J Proteomics 2019 Jun 30;202:103370.
    pmc: PMC6549134pubmed: 31028944doi: 10.1016/j.jprot.2019.04.020google scholar: lookup
  6. Madsen R, Lundstedt T, Trygg J. Chemometrics in metabolomics--a review in human disease diagnosis.. Anal Chim Acta 2010 Feb 5;659(1-2):23-33.
    pubmed: 20103103doi: 10.1016/j.aca.2009.11.042google scholar: lookup
  7. Anderson JR, Chokesuwattanaskul S, Phelan MM, Welting TJM, Lian LY, Peffers MJ, Wright HL. (1)H NMR Metabolomics Identifies Underlying Inflammatory Pathology in Osteoarthritis and Rheumatoid Arthritis Synovial Joints.. J Proteome Res 2018 Nov 2;17(11):3780-3790.
    pmc: PMC6220363pubmed: 30229649doi: 10.1021/acs.jproteome.8b00455google scholar: lookup
  8. Carlson AK, Rawle RA, Wallace CW, Brooks EG, Adams E, Greenwood MC, Olmer M, Lotz MK, Bothner B, June RK. Characterization of synovial fluid metabolomic phenotypes of cartilage morphological changes associated with osteoarthritis.. Osteoarthritis Cartilage 2019 Aug;27(8):1174-1184.
    pmc: PMC6646055pubmed: 31028882doi: 10.1016/j.joca.2019.04.007google scholar: lookup
  9. Altindag O, Erel O, Aksoy N, Selek S, Celik H, Karaoglanoglu M. Increased oxidative stress and its relation with collagen metabolism in knee osteoarthritis.. Rheumatol Int 2007 Feb;27(4):339-44.
    pubmed: 17096092doi: 10.1007/s00296-006-0247-8google scholar: lookup
  10. de Grauw JC, van de Lest CH, van Weeren PR. A targeted lipidomics approach to the study of eicosanoid release in synovial joints.. Arthritis Res Ther 2011 Jul 27;13(4):R123.
    pmc: PMC3239362pubmed: 21794148doi: 10.1186/ar3427google scholar: lookup
  11. Anderson JR, Phelan MM, Clegg PD, Peffers MJ, Rubio-Martinez LM. Synovial Fluid Metabolites Differentiate between Septic and Nonseptic Joint Pathologies.. J Proteome Res 2018 Aug 3;17(8):2735-2743.
    pmc: PMC6092013pubmed: 29969035doi: 10.1021/acs.jproteome.8b00190google scholar: lookup
  12. Graham RJTY, Anderson JR, Phelan MM, Cillan-Garcia E, Bladon BM, Taylor SE. Metabolomic analysis of synovial fluid from Thoroughbred racehorses diagnosed with palmar osteochondral disease using magnetic resonance imaging.. Equine Vet J 2020 May;52(3):384-390.
    pubmed: 31657070doi: 10.1111/evj.13199google scholar: lookup
  13. Silsirivanit A. Glycosylation markers in cancer.. Adv Clin Chem 2019;89:189-213.
    pubmed: 30797469doi: 10.1016/bs.acc.2018.12.005google scholar: lookup
  14. Li X, Xu J, Li M, Zeng X, Wang J, Hu C. Aberrant glycosylation in autoimmune disease.. Clin Exp Rheumatol 2020 Jul-Aug;38(4):767-775.
    pubmed: 31694739
  15. Alter G, Ottenhoff THM, Joosten SA. Antibody glycosylation in inflammation, disease and vaccination.. Semin Immunol 2018 Oct;39:102-110.
    pmc: PMC8731230pubmed: 29903548doi: 10.1016/j.smim.2018.05.003google scholar: lookup
  16. Pilobello KT, Krishnamoorthy L, Slawek D, Mahal LK. Development of a lectin microarray for the rapid analysis of protein glycopatterns.. Chembiochem 2005 Jun;6(6):985-9.
    pubmed: 15798991doi: 10.1002/cbic.200400403google scholar: lookup
  17. Pilobello KT, Slawek DE, Mahal LK. A ratiometric lectin microarray approach to analysis of the dynamic mammalian glycome.. Proc Natl Acad Sci U S A 2007 Jul 10;104(28):11534-9.
    pmc: PMC1913879pubmed: 17606908doi: 10.1073/pnas.0704954104google scholar: lookup
  18. Wagatsuma T, Nagai-Okatani C, Matsuda A, Masugi Y, Imaoka M, Yamazaki K, Sakamoto M, Kuno A. Discovery of Pancreatic Ductal Adenocarcinoma-Related Aberrant Glycosylations: A Multilateral Approach of Lectin Microarray-Based Tissue Glycomic Profiling With Public Transcriptomic Datasets.. Front Oncol 2020;10:338.
    pmc: PMC7082313pubmed: 32232009doi: 10.3389/fonc.2020.00338google scholar: lookup
  19. Takeshita M, Kuno A, Suzuki K, Matsuda A, Shimazaki H, Nakagawa T, Otomo Y, Kabe Y, Suematsu M, Narimatsu H, Takeuchi T. Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis.. Arthritis Res Ther 2016 May 21;18(1):112.
    pmc: PMC4875599pubmed: 27209430doi: 10.1186/s13075-016-1013-2google scholar: lookup
  20. Liang Y, Han P, Wang T, Ren H, Gao L, Shi P, Zhang S, Yang A, Li Z, Chen M. Stage-associated differences in the serum N- and O-glycan profiles of patients with non-small cell lung cancer.. Clin Proteomics 2019;16:20.
    pmc: PMC6509814pubmed: 31168300doi: 10.1186/s12014-019-9240-6google scholar: lookup
  21. Svala E, Jin C, Rüetschi U, Ekman S, Lindahl A, Karlsson NG, Skiöldebrand E. Characterisation of lubricin in synovial fluid from horses with osteoarthritis.. Equine Vet J 2017 Jan;49(1):116-123.
    pubmed: 26507102doi: 10.1111/evj.12521google scholar: lookup
  22. Estrella RP, Whitelock JM, Packer NH, Karlsson NG. The glycosylation of human synovial lubricin: implications for its role in inflammation.. Biochem J 2010 Jul 15;429(2):359-67.
    pubmed: 20443780doi: 10.1042/bj20100360google scholar: lookup
  23. Kamm JL, Nixon AJ, Witte TH. Cytokine and catabolic enzyme expression in synovium, synovial fluid and articular cartilage of naturally osteoarthritic equine carpi.. Equine Vet J 2010 Nov;42(8):693-9.
    pmc: PMC4183755pubmed: 21039798doi: 10.1111/j.2042-3306.2010.00140.xgoogle scholar: lookup
  24. Reesink HL, Watts AE, Mohammed HO, Jay GD, Nixon AJ. Lubricin/proteoglycan 4 increases in both experimental and naturally occurring equine osteoarthritis.. Osteoarthritis Cartilage 2017 Jan;25(1):128-137.
    pmc: PMC5489058pubmed: 27498214doi: 10.1016/j.joca.2016.07.021google scholar: lookup
  25. Pilobello KT, Agrawal P, Rouse R, Mahal LK. Advances in lectin microarray technology: optimized protocols for piezoelectric print conditions.. Curr Protoc Chem Biol 2013;5(1):1-23.
    pmc: PMC4734107pubmed: 23788322doi: 10.1002/9780470559277.ch120035google scholar: lookup
  26. Koppolu S, Wang L, Mathur A, Nigam JA, Dezzutti CS, Isaacs C, Meyn L, Bunge KE, Moncla BJ, Hillier SL, Rohan LC, Mahal LK. Vaginal Product Formulation Alters the Innate Antiviral Activity and Glycome of Cervicovaginal Fluids with Implications for Viral Susceptibility.. ACS Infect Dis 2018 Nov 9;4(11):1613-1622.
    doi: 10.1021/acsinfecdis.8b00157pubmed: 30183260google scholar: lookup
  27. Anderson DD, Chubinskaya S, Guilak F, Martin JA, Oegema TR, Olson SA, Buckwalter JA. Post-traumatic osteoarthritis: improved understanding and opportunities for early intervention.. J Orthop Res 2011 Jun;29(6):802-9.
    doi: 10.1002/jor.21359pmc: PMC3082940pubmed: 21520254google scholar: lookup
  28. Toegel S, Bieder D, André S, Altmann F, Walzer SM, Kaltner H, Hofstaetter JG, Windhager R, Gabius HJ. Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein.. Arthritis Res Ther 2013 Oct 4;15(5):R147.
    doi: 10.1186/ar4330pmc: PMC3978707pubmed: 24289744google scholar: lookup
  29. Gao N, Bergstrom K, Fu J, Xie B, Chen W, Xia L. Loss of intestinal O-glycans promotes spontaneous duodenal tumors.. Am J Physiol Gastrointest Liver Physiol 2016 Jul 1;311(1):G74-83.
    doi: 10.1152/ajpgi.00060.2016pmc: PMC4967177pubmed: 27229122google scholar: lookup
  30. Peal BT, Gagliardi R, Su J, Fortier LA, Delco ML, Nixon AJ, Reesink HL. Synovial fluid lubricin and hyaluronan are altered in equine osteochondral fragmentation, cartilage impact injury, and full-thickness cartilage defect models.. J Orthop Res 2020 Aug;38(8):1826-1835.
    doi: 10.1002/jor.24597pmc: PMC7354223pubmed: 31965593google scholar: lookup
  31. Ballard BL, Antonacci JM, Temple-Wong MM, Hui AY, Schumacher BL, Bugbee WD, Schwartz AK, Girard PJ, Sah RL. Effect of tibial plateau fracture on lubrication function and composition of synovial fluid.. J Bone Joint Surg Am 2012 May 16;94(10):e64.
    doi: 10.2106/jbjs.k.00046pmc: PMC3349914pubmed: 22617930google scholar: lookup
  32. Neu CP, Reddi AH, Komvopoulos K, Schmid TM, Di Cesare PE. Increased friction coefficient and superficial zone protein expression in patients with advanced osteoarthritis.. Arthritis Rheum 2010 Sep;62(9):2680-7.
    pmc: PMC2946421pubmed: 20499384doi: 10.1002/art.27577google scholar: lookup
  33. Ali L, Flowers SA, Jin C, Bennet EP, Ekwall AK, Karlsson NG. The O-glycomap of lubricin, a novel mucin responsible for joint lubrication, identified by site-specific glycopeptide analysis.. Mol Cell Proteomics 2014 Dec;13(12):3396-409.
    doi: 10.1074/mcp.m114.040865pmc: PMC4256492pubmed: 25187573google scholar: lookup
  34. Carlson AK, Rawle RA, Wallace CW, Adams E, Greenwood MC, Bothner B, June RK. Global metabolomic profiling of human synovial fluid for rheumatoid arthritis biomarkers.. Clin Exp Rheumatol 2019 May-Jun;37(3):393-399.
    pubmed: 30620276
  35. Haudenschild DR, Carlson AK, Zignego DL, Yik JHN, Hilmer JK, June RK. Inhibition of early response genes prevents changes in global joint metabolomic profiles in mouse post-traumatic osteoarthritis.. Osteoarthritis Cartilage 2019 Mar;27(3):504-512.
    pmc: PMC6391201pubmed: 30572121doi: 10.1016/j.joca.2018.11.006google scholar: lookup
  36. FLINN JH, PRICE JM, YESS N, BROWN RR. EXCRETION OF TRYPTOPHAN METABOLITES BY PATIENTS WITH RHEUMATOID ARTHRITIS.. Arthritis Rheum 1964 Jun;7:201-10.
    pubmed: 14168449doi: 10.1002/art.1780070303google scholar: lookup
  37. Tetlow LC, Woolley DE. Histamine stimulates the proliferation of human articular chondrocytes in vitro and is expressed by chondrocytes in osteoarthritic cartilage.. Ann Rheum Dis 2003 Oct;62(10):991-4.
    pmc: PMC1754325pubmed: 12972479doi: 10.1136/ard.62.10.991google scholar: lookup
  38. Carlson AK, Rawle RA, Wallace CW, Brooks EG, Adams E, Greenwood MC. Characterization of osteoarthritis phenotypes by global metabolomic profiling of human synovial fluid. bioRxiv 2018.
    doi: 10.1101/395020google scholar: lookup
  39. Rath M, Müller I, Kropf P, Closs EI, Munder M. Metabolism via Arginase or Nitric Oxide Synthase: Two Competing Arginine Pathways in Macrophages.. Front Immunol 2014;5:532.
    pmc: PMC4209874pubmed: 25386178doi: 10.3389/fimmu.2014.00532google scholar: lookup
  40. Zhu S, Zhu J, Xiao J, Ren L, Liu L, Zhou Y. Long-time fulvic acid supplementation modulates hydroxylysyl glycosylation of collagen in mice.. J Huazhong Univ Sci Technolog Med Sci 2004;24(5):427-9.
    pubmed: 15641683doi: 10.1007/bf02831099google scholar: lookup
  41. Williams RO. Collagen-induced arthritis in mice: a major role for tumor necrosis factor-alpha.. Methods Mol Biol 2007;361:265-84.
    pubmed: 17172717doi: 10.1385/1-59745-208-4:265google scholar: lookup
  42. Crowley DC, Lau FC, Sharma P, Evans M, Guthrie N, Bagchi M, Bagchi D, Dey DK, Raychaudhuri SP. Safety and efficacy of undenatured type II collagen in the treatment of osteoarthritis of the knee: a clinical trial.. Int J Med Sci 2009 Oct 9;6(6):312-21.
    pmc: PMC2764342pubmed: 19847319doi: 10.7150/ijms.6.312google scholar: lookup
  43. Klein DJ, McKeever KH, Mirek ET, Anthony TG. Metabolomic Response of Equine Skeletal Muscle to Acute Fatiguing Exercise and Training.. Front Physiol 2020;11:110.
    pmc: PMC7040365pubmed: 32132934doi: 10.3389/fphys.2020.00110google scholar: lookup
  44. Walter J, Huwiler F, Fortes C, Grossmann J, Roschitzki B, Hu J, Naegeli H, Laczko E, Bleul U. Analysis of the equine "cumulome" reveals major metabolic aberrations after maturation in vitro.. BMC Genomics 2019 Jul 17;20(1):588.
    pmc: PMC6637639pubmed: 31315563doi: 10.1186/s12864-019-5836-5google scholar: lookup

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