EV-associated miRNAs from pleural lavage as potential diagnostic biomarkers in lung cancer

Berta Roman-Canal,Julia Ponomarenko,Eva Colas,José Luis Recuero,Antonio Gil-Moreno,Maria Ruiz-Miró,Cristian Pablo Moiola,Sarah Bonnin,Esperanza González,Amaia Ojanguren,José M Porcel,Juan M Falcón-Pérez,Xavier Matias-Guiu,Sònia Gatius

doi:10.1038/s41598-019-51578-y

Abstract

Lung cancer is the leading cause of cancer-related deaths among men and women in the world, accounting for the 25% of cancer mortality. Early diagnosis is an unmet clinical issue. In this work, we focused to develop a novel approach to identify highly sensitive and specific biomarkers by investigating the use of extracellular vesicles (EVs) isolated from the pleural lavage, a proximal fluid in lung cancer patients, as a source of potential biomarkers. We isolated EVs by ultracentrifuge method from 25 control pleural fluids and 21 pleural lavages from lung cancer patients. Analysis of the expression of EV-associated miRNAs was performed using Taqman OpenArray technology through which we could detect 288 out of the 754 miRNAs that were contained in the OpenArray. The differential expression analysis yielded a list of 14 miRNAs that were significantly dysregulated (adj. p-value < 0.05 and logFC lower or higher than 3). Using Machine Learning approach we discovered the lung cancer diagnostic biomarkers; miRNA-1-3p, miRNA-144-5p and miRNA-150-5p were found to be the best by accuracy. Accordance with our finding, these miRNAs have been related to cancer processes in previous studies. This results opens the avenue to the use of EV-associated miRNA of pleural fluids and lavages as an untapped source of biomarkers, and specifically, identifies miRNA-1-3p, miRNA-144-5p and miRNA 150-5p as promising biomarkers of lung cancer diagnosis.

Highlights

Lung cancer is the leading cause of cancer-related deaths among men and women in the world, accounting for the 25% of cancer mortality
Several studies have shown the suitability of pleural lavage cytology in early stage surgically resected non-small cell lung cancer (NSCLC)
MiRNAs are a highly conserved family of small, non-coding RNAs, 19–24 nucleotides in length. They negatively regulate the expression of multiple genes either by including translational silencing or by causing the degradation of messenger RNAs of the targeted gene, via incomplete base-pairing to a complementary sequence in the 3′-untranslated region (UTR)[6]

Summary

Introduction

Lung cancer is the leading cause of cancer-related deaths among men and women in the world, accounting for the 25% of cancer mortality. Accordance with our finding, these miRNAs have been related to cancer processes in previous studies This results opens the avenue to the use of EV-associated miRNA of pleural fluids and lavages as an untapped source of biomarkers, and identifies miRNA-1-3p, miRNA-144-5p and miRNA 150-5p as promising biomarkers of lung cancer diagnosis. Detection of tumor cells in pleural lavage of patients with early stage NSCLC has shown to be associated with shorter overall survival[4]. EVs are 20–200 nm round membrane vesicles released by multivesicular bodies fusing with the cell membrane Their principal function is to participate in the intercellular communication and because of their content in bioactive material such proteins, metabolites, RNA and miRNAs, EVs have been considered an important source of biomarkers for the scientific community. This material is well-protected owing to the EVs lipid bilayer membrane, even if EVs are extracted from circulating or proximal body fluids[7]

Methods

Results

Conclusion