TY - JOUR T1 - Extended lifespan of bronchial epithelial cells maintains normal cellular phenotype and transcriptome integrity JF - ERJ Open Research JO - erjor DO - 10.1183/23120541.00254-2020 VL - 7 IS - 1 SP - 00254-2020 AU - Jonathan O'Loughlin AU - Robert J. Hall AU - Sangita Bhaker AU - Michael A. Portelli AU - Amanda Henry AU - Vincent Pang AU - David O. Bates AU - Tyson V. Sharp AU - Ian Sayers Y1 - 2021/01/01 UR - http://openres.ersjournals.com/content/7/1/00254-2020.abstract N2 - Genetic studies have identified several epithelial-derived genes associated with airway diseases. However, techniques used to study gene function frequently exceed the proliferative potential of primary human bronchial epithelial cells (HBECs) isolated from patients. Increased expression of the polycomb group protein BMI-1 extends the lifespan of HBECs while maintaining cell context plasticity. Herein we aimed to assess how BMI-1 expression impacted cellular functions and global mRNA expression. HBECs from six donors were transduced with lentivirus containing BMI-1 and cells were characterised, including by RNA sequencing and impedance measurement. BMI-1-expressing HBECs (B-HBECs) have a proliferative advantage and show comparable in vitro properties to low passage primary HBECs, including cell attachment/spreading and barrier formation. The B-HBEC mRNA signature was modestly different to HBECs, with only 293 genes differentially expressed (5% false discovery rate). Genes linked to epithelial mesenchymal transition and cell cycle were enriched in B-HBECs. We investigated the expression of genes implicated in asthma from genetic and expression studies and found that 97.6% of genes remained unaltered. We have shown that increased BMI-1 expression in HBECs delays lung epithelial cell senescence by promoting cell cycle progression and highlighted the flexible utility for B-HBECs as an important platform for studying airway epithelial mechanisms.A method to extend the lifespan of primary human bronchial epithelial cells that maintain a normal epithelial cell phenotype, thus providing a platform to investigate respiratory disease mechanisms over prolonged periods https://bit.ly/353Rklc ER -