Cigarette smoke modulates cellular responses of differentiated human bronchial epithelial cells to rhinovirus-A16 infection

Rhinovirus (RV) is one of common detected viruses during viral COPD exacerbations. We aim to study the effect of the main risk factor for COPD, cigarette smoke (CS), on RV-A16-infected well-differentiated bronchial epithelial cell cultures obtained from COPD and non-COPD patients. Primary bronchial epithelial cells (PBEC) were obtained from macroscopically normal, resected bronchial tissue from COPD and non-COPD patients (n=7/6) undergoing resection surgery for lung cancer. Well-differentiated PBEC were exposed to whole CS (WCS) for 15 minutes or air and then infected with RV-A16 (MOI 1) or mock for 1 hour. Bulk RNA sequencing (RNAseq) was performed on cells collected at 6 and 24 hours post infection (hpi). WCS exposure increased viral replication at 24 and 48 hpi compared to air controls with no significant difference between COPD and non-COPD donors. In both COPD and non-COPD groups, WCS-exposed PBEC of 6 hpi had a reduction in anti-viral responses and an increase in genes related to autophagy, canonical WNT-signaling and oxidative stress compared to air controls. At 24 hpi, in both groups, expression of genes relating to the interferon response, necroptosis and complement pathway was increased in WCS-exposed RV-A16-infected PBEC compared to air exposed RV-A16-infected PBEC. Glycolysis, oxidative phosphorylation and necroptosis-related genes were significantly higher in WCS-exposed RV-A16-infected PBEC in COPD donors compared to non-COPD donors. Overall, this study identifies specific pathways that are altered by (combined) CS and RV-A16 exposure, highly relevant for improved understanding of virus-induced COPD exacerbations. -Funded by Boehringer Ingelheim.