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Journal of proteome research2020; 19(9); 3652-3667; doi: 10.1021/acs.jproteome.0c00143

Ex Vivo Equine Cartilage Explant Osteoarthritis Model: A Metabolomics and Proteomics Study.

Abstract: Osteoarthritis is an age-related degenerative musculoskeletal disease characterized by loss of articular cartilage, synovitis, and subchondral bone sclerosis. Osteoarthritis pathogenesis is yet to be fully elucidated with no osteoarthritis-specific biomarkers in clinical use. equine cartilage explants ( = 5) were incubated in tumor necrosis factor-α (TNF-α)/interleukin-1β (IL-1β)-supplemented culture media for 8 days, with the media removed and replaced at 2, 5, and 8 days. Acetonitrile metabolite extractions of 8 day cartilage explants and media samples at all time points underwent one-dimensional (1D) H nuclear magnetic resonance metabolomic analysis, with media samples also undergoing mass spectrometry proteomic analysis. Within the cartilage, glucose and lysine were elevated following TNF-α/IL-1β treatment, while adenosine, alanine, betaine, creatine, myo-inositol, and uridine decreased. Within the culture media, 4, 4, and 6 differentially abundant metabolites and 154, 138, and 72 differentially abundant proteins were identified at 1-2, 3-5, and 6-8 days, respectively, including reduced alanine and increased isoleucine, enolase 1, vimentin, and lamin A/C following treatment. Nine potential novel osteoarthritis neopeptides were elevated in the treated media. Implicated pathways were dominated by those involved in cellular movement. Our innovative study has provided insightful information on early osteoarthritis pathogenesis, enabling potential translation for clinical markers and possible new therapeutic targets.
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Publication Date: 2020-08-06 PubMed ID: 32701294PubMed Central: PMC7476031DOI: 10.1021/acs.jproteome.0c00143Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study explores the cause of osteoarthritis, a degenerative disease that results in joint damage, by using an ex vivo equine cartilage explant and examining the metabolic and proteomic changes that occur when treated with specific inflammatory proteins. The analysis identifies several changes that could potentially be used as therapeutic targets and biomarkers for osteoarthritis.

Research Context

  • Osteoarthritis (OA) is a degenerative musculoskeletal disorder typically associated with aging. It involves the loss of cartilage that cushions joints, inflammation of the synovium or joint lining, and hardening of the subchondral bone beneath the cartilage.
  • Despite the impact of OA on the quality of life for many aging individuals, our understanding of its underlying mechanisms is still incomplete. This has resulted in a lack of biomarkers specifically associated with OA that could be used in clinical settings, thereby hindering early diagnosis and effective treatment.

Research Method

  • In an attempt to better understand OA and identify potential biomarkers, the researchers utilized ex vivo equine cartilage explants. This means that they used horse cartilage tissue which was studied outside of the living organism in a controlled environment.
  • This tissue was incubated in a media containing two pro-inflammatory proteins, tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), thought to be involved in OA.
  • The researchers then examined the metabolites (small molecules involved in metabolic processes) in the cartilage tissue and in the media in which it was cultured. Changes in these metabolites could indicate metabolic processes influencing the development of OA.
  • Protein analysis was also performed on the media samples to track any changes in protein abundance that could be linked to OA pathogenesis.

Research Findings

  • The metabolite analysis revealed changes in the levels of several substances within the cartilage tissue following the treatment with TNF-α/IL-1β. Some, such as glucose and lysine, were increased, while others, such as adenosine, alanine, and uridine, were decreased.
  • In the media, observations of differing levels of metabolites and proteins over three different time intervals indicated metabolic and protein-based changes could be occurring in response to the treatment. This included a decrease in alanine and an increase in isoleucine, enolase 1, vimentin, and lamin A/C.
  • The authors also identified nine potential osteoarthritis-related neopeptides (small protein-like molecules) increased in the treated media.
  • Much of the changes identified suggest involvement of cellular movement-related pathways in OA progression.

Research Implications

  • Overall, this research has added valuable insights into the early developments of OA at the molecular level. It presents a potential translation of these findings into clinical markers for early detection and therapeutic targets.
  • The findings also suggest new directions for future research, including looking into the potential of the identified metabolites, proteins and neopeptides as biomarkers or therapeutic targets, and further probing the role of cellular movement in OA progression.

Cite This Article

APA
Anderson JR, Phelan MM, Foddy L, Clegg PD, Peffers MJ. (2020). Ex Vivo Equine Cartilage Explant Osteoarthritis Model: A Metabolomics and Proteomics Study. J Proteome Res, 19(9), 3652-3667. https://doi.org/10.1021/acs.jproteome.0c00143

Publication

Journal of proteome research
ISSN: 1535-3907
NlmUniqueID: 101128775
Country: United States
Language: English
Volume: 19
Issue: 9
Pages: 3652-3667

Researcher Affiliations

Anderson, James R
  • Musculoskeletal and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, U.K.
Phelan, Marie M
  • NMR Metabolomics Facility, Technology Directorate & Department of Biochemistry & Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool L69 7ZB, U.K.
Foddy, Laura
  • School of Veterinary Science, Institute of Infection, Veterinary & Ecological Sciences, University of Liverpool, Liverpool L69 3GH, U.K.
Clegg, Peter D
  • Musculoskeletal and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, U.K.
Peffers, Mandy J
  • Musculoskeletal and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, U.K.

MeSH Terms

  • Animals
  • Cartilage, Articular
  • Horses
  • Interleukin-1beta
  • Metabolomics
  • Osteoarthritis
  • Proteomics
  • Tumor Necrosis Factor-alpha

Grant Funding

  • MR/P020941/1 / Medical Research Council
  • MR/M009114/1 / Medical Research Council
  • Versus Arthritis
  • 107471/Z/15/Z / Wellcome Trust
  • MR/R502182/1 / Medical Research Council

Conflict of Interest Statement

The authors declare no competing financial interest.

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