Metabolomic predictors of non-response to methotrexate therapy in patients with rheumatoid arthritis

Aim. To identify a group of metabolites potentially characterizing the lack of response to initiated therapy with disease-modifying antirheumatic drugs (methotrexate) after 24 weeks, based on an analysis of the relationship between activity parameters and the baseline metabolic profile.

Material and methods. The study group included 37 patients with RA [age 57.51 [52.63–62.40] years, 12 men (32.4 %)] who were initiated on methotrexate (MTX) therapy; the control group included 31 healthy volunteers [age 41.00 [30.00–44.00] years, 11 men (35.5 %)]. Plasma metabolites of the study participants were analyzed using ultra-high-performance liquid chromatography combined with tandem mass spectrometry. Metabolomic profiling of patients in both groups was performed at the time of study inclusion. After 24 weeks of MTX therapy, patients in the main group underwent a follow-up examination to assess treatment efficacy. Treatment response was defined as achieving low disease activity or clinical remission based on the Disease Activity Score (DAS 28-CRP). Based on this, patients in the main group were divided into «responders» and «non-responders».

Results. Metabolite classes that statistically differed significantly between RA patients and healthy volunteers included branched-chain amino acids, tryptophan metabolites, fatty acids, and urea cycle metabolites. After 24 weeks of MTX therapy, 16 patients achieved remission/low activity based on the DAS 28-CRP scale, and 21 patients achieved moderate/high activity. Moreover, patients who responded to MTX treatment had statistically higher DAS 28-CRP activity levels at baseline compared to patients who did not respond to MTX therapy (4.80 [4.40–5.45] vs. 3.99 [3.32–4.98], p=0.006). At baseline, patients in the «responder» and «non-responder» groups were comparable in age (56.27±16.58 and 54.91±11.03 years, respectively, p=0.766) and gender (4 (25.0 %) and 5 (22.7 %) men, respectively, p=1.000). However, among patients who did not respond to MTX therapy, there were more patients seropositive for both rheumatoid factor (RF) [16 (76.2 %) vs. 10 (62.5 %), p<0.05] and anti-citrullinated protein antibodies (ACP) [16 (76.2 %) vs. 8 (50.0 %), p<0.05], and the ACP level was statistically significantly higher compared to the group of patients who responded to therapy (20.00 [10.00; 422.50] vs. 400.00 [100.00; 520.00] U/ml, p=0.001). Using principal component analysis with a reliable assessment of the VIP-score (variable importance in projection), a panel of biomarkers predicting potential lack of response to MTX therapy was generated. The panel included aspartate (p<0.05), glutamate (p<0.05), betaine (p<0.05), dimethylglycine (p<0.05), tryptophan (p<0.05), quinolinic acid (p<0.05), octanoyl-carnitine (p<0.05), decanoyl-carnitine (p<0.05), asymmetric dimethylarginine (p<0.05), and uridine (p<0.05).

Conclusion. The study of the metabolomic profile of patients with RA demonstrates the potential for predicting therapy response, which in the future will serve as the basis for the development of more effective therapeutic strategies and the creation of targeted therapies aimed at metabolic processes.

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