O'Loughlin Jonathan, Hall Robert J, Bhaker Sangita, Portelli Michael A, Henry Amanda, Pang Vincent, Bates David O, Sharp Tyson V, Sayers Ian
Division of Respiratory Medicine, National Institute for Health Research, Nottingham Biomedical Research Centre, Biodiscovery Institute, University of Nottingham, Nottingham, UK.
These authors contributed equally.
ERJ Open Res. 2021 Jan 18;7(1). doi: 10.1183/23120541.00254-2020. eCollection 2021 Jan.
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 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.
基因研究已经确定了几个与气道疾病相关的上皮来源基因。然而，用于研究基因功能的技术常常超出从患者分离的原代人支气管上皮细胞（HBECs）的增殖潜能。多梳蛋白组蛋白BMI-1表达的增加延长了HBECs的寿命，同时保持细胞背景可塑性。在此，我们旨在评估BMI-1表达如何影响细胞功能和整体mRNA表达。用含有BMI-1的慢病毒转导来自六个供体的HBECs，并对细胞进行表征，包括通过RNA测序和阻抗测量。表达BMI-1的HBECs（B-HBECs）具有增殖优势，并且表现出与低代原代HBECs相当的特性，包括细胞附着/铺展和屏障形成。B-HBECs的mRNA特征与HBECs略有不同，只有293个基因差异表达（错误发现率为5%）。与上皮-间质转化和细胞周期相关的基因在B-HBECs中富集。我们研究了遗传和表达研究中涉及哮喘的基因表达，发现97.6%的基因保持不变。我们已经表明，HBECs中BMI-1表达的增加通过促进细胞周期进程延迟肺上皮细胞衰老，并强调了B-HBECs作为研究气道上皮机制的重要平台的灵活实用性。
