Genome-Wide Association Study: Functional Variant rs2076295 Regulates Desmoplakin Expression in Airway Epithelial Cells.

Genetic association studies have identified rs2076295 in association with idiopathic pulmonary fibrosis (IPF). We hypothesized that rs2076295 is the functional variant regulating () expression in human bronchial epithelial cells, and regulates extracellular matrix-related gene expression and cell migration, which is relevant to IPF development. To determine whether rs2076295 regulates expression and the function of DSP in airway epithelial cells. Using CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 editing (including regional deletion, indel, CRISPR interference, and single-base editing), we modified rs2076295 and measured expression in edited 16HBE14o- and primary airway epithelial cells. Cellular integrity, migration, and genome-wide gene expression changes were examined in 16HBE14o- single colonies with knockout. The expression of and its relevant matrix genes was measured by quantitative PCR and also analyzed in single-cell RNA-sequencing data from control and IPF lungs. is expressed predominantly in bronchial and alveolar epithelial cells, with reduced expression in alveolar epithelial cells in IPF lungs. The deletion of the DNA region-spanning rs2076295 led to reduced expression of , and the edited rs2076295GG 16HBE14o- line has lower expression of than the rs2076295TT lines. Knockout of in 16HBE14o- cells decreased transepithelial resistance but increased cell migration, with increased expression of extracellular matrix-related genes, including and . Silencing of and abolished increased migration in -knockout cells. rs2076295 regulates expression in human airway epithelial cells. The loss of enhances extracellular matrix-related gene expression and promotes cell migration, which may contribute to the pathogenesis of IPF.