Serum Mir-4254, Mir-19a and Mir-33b Are Potential Markers For Diagnosis and Prognostic Evaluation In Multiple Myeloma

A sensitive, noninvasive diagnosis and better disease classification would be very useful for improve the outcome of multiple myeloma. In this study, we proposed a microarray based biology approach to identify miRNAs as new biomarkers for the diagnosis and prognostic evaluation of multiple myeloma.

Microarray-based miRNAs expression profiling data indicated that 95 mature miRNAs were differentially expressed between myeloma patients (n=7) and healthy subjects (n=5, fold change>3.0 with P<0.01). The decrease of miR-19a/miR-92a (P<0.0001) and the increase of miR-214, 135b, 4254, 3658 and 33b (P<0.05) in myeloma patients were further validation confirmed by Q-PCR. The expression pattern of these miRNAs (miR-19a, miR-92a, miR-214, miR-135b, miR-3658, miR-4254 and miR-33b) were further detected by Taqman probes Q-PCR in 84 newly diagnosed patients, 34 cases in remission, 40 relapsed patients and 34 healthy controls. The data showed the expression of miR-19a (1.39±0.33 vs 2.56±0.18) and miR-92a (0.35±0.20 vs 1.73±0.13) were significantly lower in newly diagnosed patients compared with healthy control subjects (all P<0.05).Whereas, miR-214 (1.68±0.12 vs 1.08±0.10), miR-135b (2.25±0.28 vs 0.01±0.05), miR-4254 (1.11±0.09 vs -0.21±0.16), miR-3658 (2.26±0.29 vs 0.47±0.09) and miR-33b (2.71±0.24 vs 2.03±0.39) were significantly higher in patients than the healthy subjects (all P<0.001). Among these miRNAs, the expression of miR-19a, miR-4254 and miR-33b were closely associated with disease progressions. The expression of miR-19a in healthy control subjects and patients in remission were significantly higher than that in newly diagnosed and relapsed patients. While miR-4254 and miR-33b were significantly lower in healthy subjects and CR patients compared to newly diagnosed and relapsed patients.

ROC (receiver operating characteristic) analysis was conducted in this study and the results showed that serum miR-4254 yielded an AUC (area under the ROC curve) of 0.9261 (P<0.001) with 79.3% sensitivity and 98.5% specificity for discriminating myeloma patients from healthy controls at a cut-off value of 0.66 (RQ value). Therefore, use of miR-4254 alone provides an excellent discrimination between healthy subjects and myeloma patients. Similarly, serum miR-19a is also a potential marker with an AUC of 0.722 ( P<0.001). At a cut-off value of 2.08 (RQ value), the sensitivity for miR-19a was 77.30% and the specificity was 70.00%. However, the AUC of miR-33b was 0.63 (P<0.05). At a cutoff value of 1.015 (RQ value), the sensitivity for miR-33b was 84.10% and the specificity was 61.0%. miR-33b is not a reliable independent marker to discriminate myeloma patients from healthy controls. Then we preformed PLS-DA in various miRNA biomarkers and found that the combination of miR-4254 and miR-19a together was an even more powerful diagnostic tool for myeloma distinguishing. The ROC curve of the classifier had an AUC of 0.950 (P<0.05). miR-4254 combined with miR-33b provided the same powerful diagnostic tool for myeloma. Moreover, Kaplan-Meier survival analysis showed that patients with higher expression of miR-19a or miR-33b had significantly favorite PFS and OS than those with low expression. However, our data did not show the expression of miR-4254 correlated with favorite survival in myeloma patient.

Serum miR-4254, miR-19a and miR-33b are novel, non-invasive, sensitive and reliable biomarkers for multiple myeloma diagnosis and prognosis evaluation.

* Asterisk with author names denotes non-ASH members.

No relevant conflicts of interest to declare.