What does this research mean for the field?

A plasma extracellular vesicle-derived miRNA panel comprising miR-195-3p, miR-222-3p, and miR-127-3p can accurately differentiate malignant from benign indeterminate (Bethesda IV/Thy3f) thyroid nodules. Novelty: ClaimNovelty.NOVEL_FINDING. Consensus alignment: ConsensusAlignment.NEUTRAL.

What question did this study set out to answer?

The research aims to identify blood-derived miRNAs as diagnostic biomarkers to differentiate malignant from benign Thy3f thyroid nodules.

May 20, 2026

Circulating extracellular Vesicles-Derived miRNAs for discriminating patients with malignant from benign Bethesda IV/Thy3f thyroid nodules

Puntos clave

The research aims to identify blood-derived miRNAs as diagnostic biomarkers to differentiate malignant from benign Thy3f thyroid nodules.
Isolated EVs from plasma of matched malignant and benign patients (n=20) via differential centrifugation.
Conducted PCR using miRCURY LNA miRNA PCR Kit for quantifying identified EV-miRNAs.
Used miR-103a-3p as a normalisation control.
Significant overexpression of miR-146b-5p, miR-222-3p, miR-127-3p, and miR-195-3p in cancer patients (p=0.0098, 0.0002, 0.0052, 0.0071, respectively).
A two-miRNA panel (miR-222-3p and miR-195-3p) achieved the highest AUC of 0.927 (p=0.0009, sensitivity 90.91%, specificity 80.00%).
Adding miR-127-3p to the panel showed a slight decrease in AUC to 0.9182 but achieved 100% specificity.

Resumen

Abstract Background Patients with indeterminate Bethesda IV/Thy3f thyroid nodules mostly undergo diagnostic hemithyroidectomy for a definitive histological diagnosis, yet only ∼20% prove malignant. Thus, there is an urgent need for diagnostic biomarkers. Blood-derived extracellular vesicles (EVs) are non-invasive liquid biopsy-based biomarkers enabling detection of cancer-driven molecular alterations. We now show that blood-derived EV-enclosed miRNAs (EV-miRNAs) can differentiate patients with malignant from benign Thy3f nodules. Methods EVs were isolated by differential centrifugation from plasma of age/gender-matched malignant and benign Thy3f nodule patients (n=20) and RNA extracted immediately using miRNAs’ Micro Kit. EV-miRNAs were identified by an omics feasibility study conducted in our lab and from the literature. PCR was performed using miRCURY LNA miRNA PCR Kit for relative quantification of EV-miRNAs. For normalisation, miR-103a-3p was used. Results Significant overexpression of plasma-derived EV-miRNA miR-146b-5p, miR-222-3p, miR-127-3p, and miR-195-3p was observed in cancer versus benign Thy3f nodule patients (p=0.0098, 0.0002, 0.0052, 0.0071, respectively). No significant difference was observed for miR-21-5p (p= 0.2213). Area under the receiver operating curve (AUC) was 0.731 for miR-146b-5p, 0.860 for miR-222-3p, 0.755 for miR-127-3p, 0.852 for miR-195-3p. A two-miRNA panel of miR-222-3p and miR-195-3p achieved the highest AUC 0.927 (p=0.0009, 95% CI= 0.810-1.000, sensitivity 90.91%, specificity 80.00% at cut-off 0.4697). Adding miR-127-3p to the panel yielded a slightly lower AUC of 0.9182 but demonstrated highest sensitivity and specificity, 90.91% and 100% respectively. Conclusion A plasma EV-miRNA panel of miR-195-3p, miR-222-3p, miR-127-3p shows strong promise as non-invasive biomarker for accurately identifying malignancy. This could improve risk stratification for Thy3f nodule, supporting clinical diagnostic applicability.

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