Sofoluwe Aderonke, Zoso Alice, Bacchetta Marc, Lemeille Sylvain, Chanson Marc
Faculty of Medicine, Department of Cell Physiology & Metabolism, University of Geneva, Geneva, Switzerland.
Faculty of Medicine, Department of Pathology & Immunology, University of Geneva, Geneva, Switzerland.
J Cyst Fibros. 2021 Jul;20(4):655-663. doi: 10.1016/j.jcf.2020.08.012. Epub 2020 Aug 29.
Cystic fibrosis (CF), a genetic disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, is characterized by dysfunction of the immune response in the airway epithelium that leads to prolonged infection, colonization and exacerbated inflammation. In this study, we determined the gene expression profile of airway epithelial cells knockdown for CFTR (CFTR KD) in response to bacterial and viral challenges.
In a first approach, polarized CFTR KD and their control counterpart (CFTR CTL) cells were stimulated with P. aeruginosa-derived virulence factor flagellin. Next, we developed a model of Influenza A virus (IAV) infection in CTL and CFTR KD polarized cells. mRNA was collected for transcriptome analysis.
Beside the expected pro-inflammatory response, Gene Set Enrichment Analysis highlighted key molecular pathways and players involved in IAV and anti-viral interferon signaling. Although IAV replication was similar in both cell types, multiplex gene expression analysis revealed changes of key immune genes dependent on time of infection that were found to be CFTR-dependent and/or IAV-dependent. Interferons are key signaling proteins/cytokines in the antibacterial and antiviral response. To evaluate their impact on the altered gene expression profile in CFTR responses to pathogens, we measured transcriptome changes after exposure to Type I-, Type II- and Type III-interferons.
Our findings reveal target genes in understanding the defective immune response in the CF airway epithelium in the context of viral infection. Information provided in this study would be useful to understand the dysfunctional immune response of the CF airway epithelium during infection.
囊性纤维化（CF）是一种由囊性纤维化跨膜传导调节因子（CFTR）基因突变引起的遗传性疾病，其特征是气道上皮免疫反应功能障碍，导致感染持续、定植和炎症加剧。在本研究中，我们确定了CFTR基因敲低（CFTR KD）的气道上皮细胞在应对细菌和病毒挑战时的基因表达谱。
在第一种方法中，用铜绿假单胞菌衍生的毒力因子鞭毛蛋白刺激极化的CFTR KD细胞及其对照细胞（CFTR CTL）。接下来，我们在CTL和CFTR KD极化细胞中建立了甲型流感病毒（IAV）感染模型。收集mRNA进行转录组分析。
除了预期的促炎反应外，基因集富集分析突出了参与IAV和抗病毒干扰素信号传导的关键分子途径和参与者。虽然IAV在两种细胞类型中的复制相似，但多重基因表达分析显示，关键免疫基因的变化取决于感染时间，且这些变化被发现与CFTR和/或IAV有关。干扰素是抗菌和抗病毒反应中的关键信号蛋白/细胞因子。为了评估它们对CFTR对病原体反应中基因表达谱改变的影响，我们测量了暴露于I型、II型和III型干扰素后的转录组变化。
我们的研究结果揭示了在病毒感染背景下理解CF气道上皮免疫反应缺陷的靶基因。本研究提供的信息将有助于理解感染期间CF气道上皮功能失调的免疫反应。
