Characterization of the MicroRNA Cargo of Extracellular Vesicles Isolated from a Pulmonary Tumor-Draining Vein Identifies miR-203a-3p as a Relapse Biomarker for Resected Non-Small Cell Lung Cancer

Abstract

In resected non-small cell lung cancer (NSCLC), post-surgical recurrence occurs in around 40% of patients, highlighting the necessity to identify relapse biomarkers. An analysis of the extracellular vesicle (EV) cargo from a pulmonary tumor-draining vein (TDV) can grant biomarker identification. We studied the pulmonary TDV EV-miRNAome to identify relapse biomarkers in a two-phase study (screening and validation). In the screening phase, a 17-miRNA relapse signature was identified in 18 selected patients by small RNAseq. The most expressed miRNA from the signature (EV-miR-203a-3p) was chosen for further validation. Pulmonary TDV EV-miR-203a-3p was studied by qRT-PCR in a validation cohort of 70 patients, where it was found to be upregulated in relapsed patients (p = 0.0194) and in patients with cancer spread to nearby lymph nodes (N+ patients) (p = 0.0396). The ROC curve analysis showed that TDV EV-miR-203a-3p was able to predict relapses with a sensitivity of 88% (AUC: 0.67; p = 0.022). Moreover, patients with high TDV EV-miR-203a-3p had a shorter time to relapse than patients with low levels (43.6 vs. 97.6 months; p = 0.00703). The multivariate analysis showed that EV-miR-203a-3p was an independent, predictive and prognostic post-surgical relapse biomarker. In conclusion, pulmonary TDV EV-miR-203a-3p is a promising new relapse biomarker for resected NSCLC patients.

A Review of Biomarkers and Their Clinical Impact in Resected Early-Stage Non-Small-Cell Lung Cancer

“…Han et al isolated several miRNAs in extracellular vesicles (EV) from pulmonary tumour drainage veins (TDVs) and identified miR-203a-3p as a recurrence marker in the 18-patient screening cohort. They further verified that miR-203a-3p was a prognostic indicator for unfavourable TTR in the 70-patient validation cohort [ 129 ].…”

MicroRNAs Present in Malignant Pleural Fluid Increase the Migration of Normal Mesothelial Cells In Vitro and May Help Discriminate between Benign and Malignant Effusions

“…It can also play a role as an oncogene by promoting drug resistance, proliferation, migration, and invasion [ 45 , 46 , 47 ]. In one study, miR-203a-3p was found to be the most highly expressed miRNA in extravesicular vesicles that were isolated from the pulmonary tumor-draining vein of patients with NSCLC, and this molecule was associated with tumor relapse [ 48 ]. Finally, miR-141-3p also plays both tumor-suppressive and tumor-supportive roles.…”

Lymphatic metastasis in non‐small cell lung cancer: recent discoveries and novel therapeutic targets

“…Additionally, EV‐miR‐378 upregulation has been reported to indicate lymphatic metastasis in NSCLC [ 19 ]. EV‐miR‐203a‐3p was also shown to be significantly increased in lymph node‐positive NSCLC patients [ 20 ]. Moreover, uregulated EV‐derived circular RNAs and long non‐coding RNAs, such as circR‐0056285 and AGAP2 antisense RNA 1 (AGAP2‐AS1), have been explored to diagnose lymphatic metastasis in NSCLC [ 21 , 22 ].…”