Changes in MiRNA-5196 Expression as a Potential Biomarker of Anti-TNF-α Therapy in Rheumatoid Arthritis and Ankylosing Spondylitis Patients

Abstract

In this study, we analysed the expression level of sera circulating miRNA-5196 in rheumatoid arthritis (RA) and ankylosing spondylitis (AS) patients before and after tumor necrosis factor (TNF)-α therapy as biomarkers predicting positive treatment outcome. We enrolled 10 RA patients, 13 AS patients, and 12 healthy individuals in the study. The expression of miRNA-5196 was measured by real-time polymerase chain reaction before and after anti-TNF-α therapy. Disease activity of RA patients was assessed using disease activity score 28 (DAS28), whereas ankylosing spondylitis DAS (ASDAS) was used in AS patients. MiRNA-5196 expression was significantly higher in patients with RA and AS before TNF-α therapy than in those following anti-TNF-α therapy and healthy controls. Changes in miRNA-5196 expression positively correlated with delta DAS28 or delta ASDAS, respectively, following TNF-α therapy. In contrast, changes in C-reactive protein (CRP) levels in RA and AS patients did not positively correlate with DAS28 or ASDAS changes. Receiver-operating characteristic analysis showed better diagnostic accuracy of miRNA-5196 expression both in RA (area under curve (AUC) = 0.87, p = 0.055) and AS patients (AUC = 0.90, p = 0.050) compared to CRP levels in RA (AUC = 0.75, p = 0.201) and AS patients (AUC = 0.85, p = 0.086) upon biologic therapy treatment. Finding novel biomarkers, including miRNA-5196 which allow to predict and monitor anti-TNF-α response, would be of clinical value especially during the early phase of RA or AS development.

Keywords: Ankylosing spondylitis; Anti-TNF-α; Biologic therapy; Biomarker; MiRNA; Rheumatoid arthritis.

Fig. 1

Level of circulating miRNA-5196 present in sera of rheumatic disease patients. The expression level of sera circulating miRNA-5196 was measured in HC (n = 15), SSc (n = 11) patients and in RA patients before (n = 10) and after (n = 10) TNF-a therapy and in AS patients before (n = 13) and after (n = 13) TNF-a therapy. Results were normalized to the let-7a internal control. Each symbol represents an individual subject; horizontal lines with bars show the mean ± SEM. P values are expressed as follows: 0.05 > P > 0.01 as *; 0.01 > P > 0.001 as **; P < 0.001 as ***

Fig. 2

Expression level of miRNA-5196, DAS28 score and CRP before and after anti-TNF-a therapy. The expression level of sera circulating miRNA-5196 (a), disease activity score DAS28 (b) and CRP (c) were measured in RA patients before (n = 10) and after (n = 10) TNF-a therapy. P values are expressed as follows: 005 > P > 001 as *; 001 > P > 0001 as **; P < 0001 as ***, ns not significant

Fig. 3

Expression level of miRNA-5196, DAS28 score and CRP before and after anti-TNF-a therapy. The expression level of sera circulating miRNA-5196 (a), ankylosing spondylitis disease activity score ASDAS (b) and CRP (c) were measured in AS patients before (n = 13) and after (n = 13) TNF-a therapy. P values are expressed as follows: 005 > P > 001 as *; 001 > P > 0001 as **; P < 0001 as ***, us not significant

Fig. 4

Correlation between changes in serum level of miRNA-5196 (delta miRNA-5196), changes in DAS28 (delta DAS28) and changes in CRP levels (delta CRP) in RA patients (n = 10) following anti-TNF-a therapy. Changes in miRNA-5196 expression were correlated with changes in DAS28 (a), changes in CRP levels were correlated with changes in DAS28 (b) and changes in CRP levels were correlated with changes in miRNA-5196 expression (c)

Fig. 5

Correlation between changes in serum level of miRNA-5196 (delta miRNA-5196), changes in ASDAS (delta ASDAS) and changes in CRP levels in AS patients (n = 9) following anti-TNF-a therapy. Changes in miRNA-5196 expression were correlated with changes in ASDAS (a), changes in CRP levels were correlated with changes in ASDAS (b) and changes in CRP levels were correlated with changes in miRNA-5196 expression (c)

Fig. 6

ROC analysis comparing delta miRNA-5196 and delta CRP levels as biomarkers predicting anti-TNF-a outcomes in RA (a) and AS patients (b). 10 RA patients and 9 AS patients were included in the analysis