Gene expression profile at single cell level of hiPSC-derived lung airways epithelial cells upon knock-out of EHF in normoxia and in hypoxia

Cystic Fibrosis (CF) is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene coding for a chloride- and bicarbonate-ion channel. Deficient or dysfunctional CFTR changes ion composition resulting in abnormal mucus, affecting transport and fluid's homeostasis in multiple epithelia. Lung disease is responsible for the majority of patients' morbimortality. Intriguingly, more than half of its phenotype is not due to CFTR mutations, but to the environment and genetic modifiers. Genome-Wide Association Studies (GWAS) have identified Single Nucleotide Polymorphisms (SNPs) that could be genetic modifiers. One of the regions outlined (11q13) is located close by an epithelium-specific gene named ETS Homologous Factor (EHF). Here, we hypothesized EHF may be impacted by these variants and may play a crucial role in the airway epithelium. We used human-induced Pluripotent Stem Cells (hiPSC)- derived Airway Epithelial Cells (AECs) to study the knock-out of EHF in normoxia and in hypoxia. Overall design: Lung airways epithelial cells were generated from hiPSC following our differentiation protocol. At the end point of differentiation, AECs seeded onto ALIs were harvested and analyzed using scRNAseq. We used isogenic background with one clone wild type and one clone homozygous for EHF knock-out.