CFTR limits F‐actin formation and promotes morphological alignment with flow in human lung microvascular endothelial cells

Abstract

Abstract Micro‐ and macrovascular endothelial dysfunction in response to shear stress has been observed in cystic fibrosis (CF), and has been associated with inflammation and oxidative stress. We tested the hypothesis that the cystic fibrosis transmembrane conductance regulator (CFTR) regulates endothelial actin cytoskeleton dynamics and cellular alignment in response to flow. Human lung microvascular endothelial cells (HLMVEC) were cultured with either the CFTR inhibitor GlyH‐101 (20 µM) or CFTRinh‐172 (20 µM), tumor necrosis factor (TNF)‐α (10 ng/ml) or a vehicle control (0.1% dimethyl sulfoxide) during 24 and 48 h of exposure to shear stress (11.1 dynes/cm2) or under static control conditions. Cellular morphology and filamentous actin (F‐actin) were assessed using immunocytochemistry. [Nitrite] and endothelin‐1 ([ET‐1]) were determined in cell culture supernatant by ozone‐based chemiluminescence and ELISA, respectively. Treatment of HLMVECs with both CFTR inhibitors prevented alignment of HLMVEC in the direction of flow after 24 and 48 h of shear stress, compared to vehicle control (both p 0.05). CFTR appears to limit cytosolic actin polymerization, while maintaining a cortical rim actin distribution that is important for maintaining barrier integrity and promoting alignment with flow, without effects on endothelial nitrite or ET‐1 production.

Keywords

• actin cytoskeleton
• cystic fibrosis
• pulmonary microvascular endothelium
• shear stress