Expression profiles of miR-101-3p and miR-431-5p as potential diagnostic biomarkers for rheumatoid arthritis

Figures

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  Fig. 1. Immunological and hematological parameters for participants with RA compared to controls. The charts show significantly higher levels (**p < 0.01) of rheumatoid factor (RF), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) at 1 h and 2 h, antinuclear antibody (ANA), and anti-cyclic citrullinated peptide (anti-CCP) antibodies in RA cases compared to controls. These parameters indicate increased inflammation and autoimmune activity in RA.

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  Fig. 2. Biochemical parameters for participants with RA compared to controls. The charts show no significant differences in SGPT, creatinine, cholesterol, triglycerides, and HbA1c levels. However, significant differences (*p < 0.05) are observed in calcium, phosphorus, and uric acid levels, with lower calcium and phosphorus and higher uric acid in RA cases, indicating altered metabolic profiles in these patients.

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  Fig. 3. Correlation coefficients between various immunological and inflammatory parameters in RA patients. The scatter plots illustrate the relationships between rheumatoid factor (RF) and C-reactive protein (CRP), erythrocyte sedimentation rate at 1 h (ESR1h), erythrocyte sedimentation rate at 2 h (ESR2h), antinuclear antibody (ANA), anti-cyclic citrullinated peptide (ANTI-CCP), and Disease Activity Score (DAS). Strong positive correlations are observed between RF and CRP, RF and ESR1h, RF and ANTI-CCP, CRP and ESR1h, CRP and ESR2h, CRP and ANTI-CCP, ESR1h and ESR2h, and DAS with RF, CRP, ESR1h, and ESR2h. A negative correlation is noted between RF and ANA. These correlations emphasize the interconnected nature of these markers in reflecting disease activity and inflammation in RA.

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  Fig. 4. ROC curves comparing the diagnostic performance of rheumatoid factor (RF), antinuclear antibody (ANA), and anti-cyclic citrullinated peptide (ANTI-CCP) for RA. The ANTI-CCP test (green line) exhibits the highest AUC of 0.965 with a sensitivity of 78% and specificity of 90%, followed by the RF test (blue line) with an AUC of 0.849, sensitivity of 65%, and specificity of 100%. The ANA test (red line) shows an AUC of 0.721 with a sensitivity of 40% and specificity of 100%. These curves indicate that ANTI-CCP is the most effective marker for RA diagnosis, followed by RF and ANA.

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  Fig. 5. Fold change in the expression levels of miR-99b-5p, miR-101-3p, and miR-431-5p in RA patients compared to healthy controls. This bar graph illustrates the relative expression levels of three different miRNAs (miR-99b-5p, miR-101-3p, and miR-431-5p) between RA patients and healthy controls. The y-axis represents the fold change in expression, indicating how much the expression levels of these miRNAs are increased or decreased in RA patients compared to the control group.

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  Fig. 6. Comparison of fold expression levels of miR-99b-5p, miR-101-3p, and miR-431-5p in RA patients undergoing conventional DMARDs treatment versus biological DMARDs treatment. miR-99b-5p (Left): Shows similar expression levels between the two treatment groups. miR-101-3p (Middle): Displays a significantly higher expression in patients treated with conventional DMARDs compared to those treated with biological DMARDs, indicated by the double asterisks (**), highlighting a significant statistical difference. miR-431-5p (Right)**: Indicates higher expression levels in the biological DMARDs treatment group compared to the conventional DMARDs group.

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  Fig. 7. Expression levels of circulating miRNAs (miR-99b-5p, miR-101-3p, and miR-431-5p) in RA patients categorized by disease duration. miR-99b-5p (Left): Expression is highest in patients with disease duration of 1–5 years, decreasing significantly in those with 5–10 years, and even more in those with over 10 years. miR-101-3p (Middle): Shows a similar trend, with the highest expression in the 1–5 years group and decreasing levels in the subsequent groups. miR-431-5p (Right): Exhibits higher expression in the 1-5- and 5-10-years groups compared to those with more than 10 years.

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  Fig. 8. Expression levels of circulating miRNAs (miR-99b-5p, miR-101-3p, and miR-431-5p) in RA patients with and without a family history of the disease. The bar graphs depict the fold expression levels of three miRNAs in RA patients, comparing those with a family history of the disease to those without. miR-99b-5p (Left): Shows similar expression levels between patients with and without a family history of RA. miR-101-3p (Middle): Displays significantly higher expression in patients with a family history of the disease compared to those without, indicated by the asterisk (*), highlighting a significant statistical difference. miR-431-5p (Right): Indicates no significant difference in expression levels between the two groups.

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  Fig. 9. Expression levels of circulating miRNAs (miR-99b-5p, miR-101-3p, and miR-431-5p) in RA patients categorized by Disease Activity Score (DAS) into remission, low, moderate, and high disease activity. miR-99b-5p (Left): Shows variable expression with higher levels in moderate and high DAS categories. miRNA-101-3p (Middle): Indicates increased expression in patients with high disease activity. miR-431-5p (Right): Exhibits elevated levels in the low DAS category compared to remission and moderate categories.

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  Fig. 10. Scatter plot with a linear regression line for the correlation between expression level of miR-101-3p and miR-431-5p. The plot demonstrates a positive correlation between the variables, as indicated by the upward slope of the regression line.

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  Fig. 11. ROC Curve Analysis for miRNAs in RA Diagnosis. The ROC curves illustrate the diagnostic performance of three miRNAs in distinguishing RA among participants. The first curve represents miR-101-3p, showing the highest Area Under the Curve (AUC) of 0.873, with a sensitivity of 78% and specificity of 94% (p-value < 0.0001). The second curve corresponds to miR-431-5p, with an AUC of 0.856, sensitivity of 73%, and specificity of 89% (p-value = 0.003). The third curve depicts miR-99b-5p, with an AUC of 0.737, sensitivity of 68%, and specificity of 89% (p-value < 0.0001). These curves demonstrate the potential of these miRNAs as reliable biomarkers for the early detection of RA.