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Epigenetic regulation of 15-lipoxygenase-1 expression in human chondrocytes by promoter methylation

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Abstract

Objective and design

15-Lipoxygenase-1 (15-LOX-1) catalyzes the biosynthesis of many anti-inflammatory and immunomodulatory lipid mediators and was reported to have protective properties in several inflammatory conditions, including osteoarthritis (OA). This study was designed to evaluate the expression of 15-LOX-1 in cartilage from normal donors and patients with OA, and to determine whether it is regulated by DNA methylation.

Methods

Cartilage samples were obtained at autopsy from normal knee joints and from OA-affected joints at the time of total knee joint replacement surgery. The expression of 15-LOX-1 was evaluated using real-time polymerase chain reaction (PCR). The role of DNA methylation in 15-LOX-1 expression was assessed using the DNA methyltransferase inhibitor 5-Aza-2′-desoxycytidine (5-Aza-dC). The effect of CpG methylation on 15-LOX-1 promoter activity was evaluated using a CpG-free luciferase vector. The DNA methylation status of the 15-LOX-1 promoter was determined by pyrosequencing.

Results

Expression of 15-LOX-1 was upregulated in OA compared to normal cartilage. Treatment with 5-Aza-dC increased 15-LOX-1 mRNA levels in chondrocytes, and in vitro methylation decreased 15-LOX-1 promoter activity. There was no difference in the methylation status of the 15-LOX-1 gene promoter between normal and OA cartilage.

Conclusion

The expression level of 15-LOX-1 was elevated in OA cartilage, which may be part of a repair process. The upregulation of 15-LOX-1 in OA cartilage was not associated with the methylation status of its promoter, suggesting that other mechanisms are involved in its upregulation.

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Data availability

All data generated during this study are included in this manuscript. The original experimental raw data can be obtained from the authors upon request.

Abbreviations

5-Aza-dC:

5-Aza-2′-desoxycytidine

15-LOX-1:

15-Lipoxygenase-1

15-HETE:

15-Hydroxyeicosatetraenoic acid

13-HODE:

13-Hydroxyoctadecadienoic acid

DNMT:

DNA methyltransferase

IL:

Interleukin

LXA4:

Lipoxin A4

OA:

Osteoarthritis

qRT-PCR:

Quantitative real-time polymerase chain reaction

RvD1:

Resolvin D1

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Funding

Canadian Institutes of Health Research (Grant: PJT-1751110 and PJT-180297). The Arthritis Society (Grant: SOG-20-0000000046).

Author information

Authors and Affiliations

  1. Osteoarthritis Research Unit, University of Montreal Hospital Research Center (CRCHUM), Montreal, QC, Canada

    Sami G. Alsabri, Gadid G. Guedi, Mehdi Najar, Jean-Pierre Pelletier, Johanne Martel-Pelletier & Hassan Fahmi

  2. LBEES, Genetics and Immune Cell Therapy Unit, Faculty of Sciences, University Mohamed Premier, Oujda, Morocco

    Makram Merimi

  3. Departement of Orthopedic Surgery, Centre Hospitalier de L’Université de Montréal (CHUM), Montréal, Québec, Canada

    Frédéric Lavoie & Julio Fernandes

  4. Department of Anatomy, Research Unit in Clinical and Functional Anatomy, Université du Québec à Trois-Rivières, Trois-Rivières, QC, Canada

    Detlev Grabs

  5. Orthopedics Research Laboratory, Research Center, Hôpital du Sacré-Cœur de Montréal, Université de Montréal, Montréal, Québec, H4J 1C5, Canada

    Julio Fernandes & Mohamed Benderdour

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  1. Sami G. Alsabri
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Contributions

SA and HF: Study conception and design. SA, GG, MN and HF: Acquisition of data. All authors contributed to the analysis and interpretation of data. All authors were also involved in manuscript writing, and all authors approved the final version.

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Correspondence to Hassan Fahmi.

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Alsabri, S.G., Guedi, G.G., Najar, M. et al. Epigenetic regulation of 15-lipoxygenase-1 expression in human chondrocytes by promoter methylation. Inflamm. Res. 72, 2145–2153 (2023). https://doi.org/10.1007/s00011-023-01805-0

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