Baseline adenosine receptor mRNA expression in blood as predictor of response to methotrexate therapy in patients with rheumatoid arthritis

Abstract

Methotrexate (MTX) reduces inflammation by increasing extracellular adenosine levels in rheumatoid arthritis (RA) patients. Adenosine acts via G-protein coupled receptors; ADORA1, ADORA2a, ADORA2b and ADORA3. We studied if baseline expression of whole blood adenosine receptors can predict response to MTX. RA patients [American College of Rheumatology/European-League-Against-Rheumatism (EULAR) 2010 criteria], Disease modifying anti-rheumatic drug (DMARD) naïve with active disease [Disease Activity Score 28 (DAS28) < 3.2] were enrolled. Blood samples were collected at baseline (n = 100) and at 4 months after therapy (n = 50). Patients were treated with MTX monotherapy. Based on EULAR response, patients were categorized into three groups i.e. good, moderate and non-responders. Adenosine receptors gene expression (ADORA1, ADORA2a, ADORA2b and ADORA3) in whole-blood RNA was measured using real-time PCR. HPRT1 was used as housekeeping gene. Receptor expression at baseline was correlated with response to MTX. All values are expressed as median (interquartile range). Hundred patients [87% females; age 40 (18) years]; duration of disease 24 (24.75) months; DAS28 4.7 (1.25) were enrolled. Fifty-one were classified as good, 28 moderate and 21 as non-responders. No expression of ADORA1 and ADORA2b was detected. Significant difference was observed in the expression levels of ADORA3 between good vs non-responder (P = 0.03) and moderate vs non-responder (P = 0.002). On ROC curve analysis, ADORA3 with cut-off value of less than - 0.60 (ΔCt) predicted non-response to MTX treatment (AUC: 0.7, P = 0.006). ADORA3 mRNA levels in whole blood may serve as a biomarker of response to MTX.