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. 2025 Mar 20:16:1499723.
doi: 10.3389/fphar.2025.1499723. eCollection 2025.

The interaction between a leflunomide-response methylation site (cg17330251) and variant (rs705379) on response to leflunomide in patients with rheumatoid arthritis

Feng Zhao  1 , Yulan Chen  1 , Haina Liu  2 , Lei Jin  3 , Xin Feng  4 , Bingbing Dai  5 , Meng Chen  1 , Qiao Wang  1 , Yuxin Yao  1 , Ruobing Liao  1 , Junyi Zhao  1 , Bingjia Qu  1 , Ying Song  1 , Lingyu Fu  1
Affiliations

The interaction between a leflunomide-response methylation site (cg17330251) and variant (rs705379) on response to leflunomide in patients with rheumatoid arthritis

Feng Zhao et al. Front Pharmacol. .

Abstract

Objectives: This research aims to reveal the mechanisms of the effect of the Paraoxonase 1 (PON1) gene on response to leflunomide (LEF) in rheumatoid arthritis (RA) patients, in terms of single nucleotide polymorphism (SNP), DNA methylation levels.

Methods: A total of 240 RA patients enrolled were categorized into the good response group and the non-response group according to the difference in DAS28 scores between baseline and 6 months after LEF administration. The identified LEF-response cytosine-phosphate-guanines (CpGs) island (cg17330251) and its internal SNPs (rs705379, etc.) located at the PON1 promoter were detected by Sanger sequencing and methyl target sequencing.

Results: A total of 12 CpG sites at cg17330251 could be identified in our RA patients. There were significant difference between the responders and non-responders in nine CpG sites: cg17330251_2, cg17330251_3, cg17330251_4, cg17330251_6, cg17330251_7, cg17330251_8, cg17330251_9, cg17330251_10, cg17330251_12, [OR (95CI%) = 0.492 (0.250, 0.969), 0.478 (0.243, 0.940), 0.492 (0.250, 0.969), 0.461 (0.234, 0.907), 0.492 (0.250, 0.969), 0.437 (0.225, 0.849), 0.478 (0.243, 0.941), 0.421 (0.212, 0.836), 0.424 (0.213, 0.843), P < 0.05, respectively]. At all these nine CpG sites, the proportions of low methylation levels in the responders were higher than those in the non-responders (P < 0.05). In a dominant model, there was a significant difference in rs705379 wildtype CC and mutant genotypes (CT + TT) between the responders and non-responders (P < 0.05). The average methylation level of 12 CpG sites was lowest in rs705379-CC (median 0.229, IQR 0.195-0.287), then rs705379-CT (median 0.363, IQR 0.332-0.395), and rs705379-TT (median:0.531, IQR:0.496-0.557). The average methylation levels of 12 CpG sites were significantly negative correlated with ΔDAS28 (r = -0.13, P < 0.05). The Logistic regression indicated that combined effect of rs705379, DNA methylation of the PON1 gene [OR (95CI%) = 1.277 [1.003, 1.626)], systemic inflammation index (SIRI) [OR (95CI%) = 1.079 (1.018, 1.143)] served as protective factors on response to LEF in RA patients.

Conclusion: The RA patients with SNP-rs705379-CC, the low methylation level of PON1-cg17330251 and more SIRI would be susceptible of response to LEF and more suitable to choose LEF treatment.

Keywords: DNA methylation4; PON1; leflunomide; rheumatoid arthritis; single nucleotide polymorphism.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Correlations among rs705379, average methylation levels of cg17330251 of PON1 and ΔDAS28. ***P < 0.001. (A) ΔDAS28 in each genotype at rs705379; (B) Average methylation level of cg17330251 in each genotype at rs705379; (C) Correlation between average methylation level of cg17330251 and ΔDAS28.
FIGURE 2
FIGURE 2
Biological interaction network of PON1. Different colors represent diverse bioinformatics methods.
FIGURE 3
FIGURE 3
GO enrichment analysis of PON1 gene.
See this image and copyright information in PMC

References

    1. Abbasi M., Mousavi M. J., Jamalzehi S., Alimohammadi R., Bezvan M. H., Mohammadi H., et al. (2019). Strategies toward rheumatoid arthritis therapy; the old and the new. J. Cell Physiol. 234 (7), 10018–10031. 10.1002/jcp.27860 - DOI - PubMed
    1. Ai R., Laragione T., Hammaker D., Boyle D. L., Wildberg A., Maeshima K., et al. (2018). Comprehensive epigenetic landscape of rheumatoid arthritis fibroblast-like synoviocytes. Nat. Commun. 9 (1), 1921. 10.1038/s41467-018-04310-9 - DOI - PMC - PubMed
    1. Aletaha D., Neogi T., Silman A. J., Funovits J., Felson D. T., Bingham C. O., 3rd, et al. (2010). 2010 Rheumatoid arthritis classification criteria: an American College of Rheumatology/European League against Rheumatism collaborative initiative. Arthritis Rheum. 62(9):2569–2581. 10.1002/art.27584 - DOI - PubMed
    1. Ardicli S., Yigitgor P., Babayev H., Ozen D., Bozkurt B., Senturk N., et al. (2024). The markers of the predictive DNA test for canine hip dysplasia may have a stronger relationship with elbow dysplasia. Heliyon 10 (18), e37716. 10.1016/j.heliyon.2024.e37716 - DOI - PMC - PubMed
    1. Arnett F. C., Edworthy S. M., Bloch D. A., McShane D. J., Fries J. F., Cooper N. S., et al. (1988). The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 31 (3), 315–324. 10.1002/art.1780310302 - DOI - PubMed

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