Effect of formoterol and tumor necrosis factor (TNFa) alone and in combination on gene expression changes in human primary bronchial epithelial cells: a comparison with forskolin treatment

RNA sequencing was performed to study the genomic effects of formoterol, a long acting ß2-adrenoceptor agonist, alone and in combination with the pro-inflammatory cytokine, TNFa in human primary bronchial epithelial cells, and to compare its effect to forskolin. Our results show that formoterol promoted a profound genomic response alone and on background of an inflammatory stimulus. Moreover, formoterol and forskolin regulated transcriptomes were highly rank order correlated which suggests that formoterol-mediated gene expression changes are cAMP dependent Overall design: Human primary bronchial epithelial cells obtained by proteinase digestion of non-transplanted, normal airways of five lung donors obtained from a tissue retrieval service at the International Institute for the Advancement of Medicine (Edison, NJ, USA), were grown as monolayers in submersion culture. Confluent cells were cultured for 1, 2, 6 and 18 h with vehicle, TNFa (10 ng ml-1), formoterol (1 nM), TNFa and formoterol in combination or forskolin (10 µM). Total RNA was extracted, quantified, processed into mRNA libraries, and sequenced across five high-throughput sequencing kits on a NextSeq 500 instrument (Illumina).