AUTHOR=Reid Laura V. , Spalluto C. Mirella , Wilkinson Tom M. A. , Staples Karl J. 

TITLE=Influenza-induced microRNA-155 expression is altered in extracellular vesicles derived from the COPD epithelium

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 15 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1560700

DOI=10.3389/fcimb.2025.1560700

ISSN=2235-2988

ABSTRACT=BackgroundInfluenza virus particularly affects those with chronic lung conditions such as Chronic Obstructive Pulmonary Disease (COPD). Airway epithelial cells are the first line of defense and primary target of influenza infection and release extracellular vesicles (EVs). EVs can transfer of biological molecules such as microRNAs (miRNAs) that can modulate the immune response to viruses through control of the innate and adaptive immune systems. The aim of this work was to profile the EV miRNAs released from bronchial epithelial cells in response to influenza infection and discover if EV miRNA expression was altered in COPD.MethodsInfluenza infection of air-liquid interface (ALI) differentiated BCi-NS1.1 epithelial cells were characterized by analyzing the expression of antiviral genes, cell barrier permeability and cell death. EVs were isolated by filtration and size exclusion chromatography from the apical surface wash of ALI cultured bronchial epithelial cells. The EV miRNA cargo was sequenced and reads mapped to miRBase. The BCi sequencing results were further investigated by RT-qPCR and by using healthy and COPD primary epithelial cells.ResultsInfection of ALI cultured BCi cells with IAV at 3.6 x 106 IU/ml for 24 h led to significant upregulation of anti-viral genes without high levels of cell death. EV release from ALI-cultured BCi cells was confirmed using electron microscopy and detection of known tetraspanin EV markers using western blot and the ExoView R100 platform. Differential expression analyses identified 5 miRNA that had a fold change of >0.6: miR-155-5p, miR-122-5p, miR-378a-3p, miR-7-5p and miR-146a-5p (FDR<0.05). Differences between EV, non-EV and cellular levels of these miRNA were detected. Primary epithelial cell release of EV and their miRNA cargo was similar to that observed for BCi. Intriguingly, miR-155 expression was decreased in EVs derived from COPD patients compared to EVs from control samples.ConclusionEpithelial EV miRNA release may be a key mechanism in modulating the response to IAV in the lungs. Furthermore, changes in EV miRNA expression may play a dysfunctional role in influenza-induced exacerbations of COPD. However, further work to fully characterize the function of EV miRNA in response to IAV in both health and COPD is required.