Mechanisms of endothelial cell dysfunction in cystic fibrosis

• Published in: Biochimica et biophysica acta. Molecular basis of disease Vol. 1863; no. 12; pp. 3243 - 3253
• Main Authors: Totani, Licia, Plebani, Roberto, Piccoli, Antonio, Di Silvestre, Sara, Lanuti, Paola, Recchiuti, Antonio, Cianci, Eleonora, Dell'Elba, Giuseppe, Sacchetti, Silvio, Patruno, Sara, Guarnieri, Simone, Mariggiò, Maria A., Mari, Veronica C., Anile, Marco, Venuta, Federico, Del Porto, Paola, Moretti, Paolo, Prioletta, Marco, Mucilli, Felice, Marchisio, Marco, Pandolfi, Assunta, Evangelista, Virgilio, Romano, Mario
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2017
• Subjects: Antigens, CD - metabolism; beta-Arrestin 2 - metabolism; Cadherins - metabolism; Cell Proliferation - physiology; Cyclic AMP; Cyclic AMP - metabolism; Cystic fibrosis; Cystic Fibrosis - metabolism; Cystic Fibrosis - pathology; Cystic Fibrosis Transmembrane Conductance Regulator - metabolism; Cytokines - metabolism; Endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - pathology; Homeostasis - physiology; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Insulin - pharmacology; Interleukin-8 - metabolism; Microvesicles; Nitric oxide; Nitric Oxide Synthase Type III - metabolism; Nitrogen Oxides - metabolism; Phosphorylation; Pulmonary Artery - metabolism; Pulmonary Artery - pathology; eNOS; YFP; PKA; Cyclic AMP; AJ; Cystic fibrosis; PAEC; Inflammation; Microvesicles; Endothelial cells; EMV; PDE; TNF-α; Nitric oxide; HUVEC; TEER; ECGF; CFTR
• ISSN: 0925-4439; 1879-260X
• DOI: 10.1016/j.bbadis.2017.08.011

Although cystic fibrosis (CF) patients exhibit signs of endothelial perturbation, the functions of the cystic fibrosis conductance regulator (CFTR) in vascular endothelial cells (EC) are poorly defined. We sought to uncover biological activities of endothelial CFTR, relevant for vascular homeostasis and inflammation. We examined cells from human umbilical cords (HUVEC) and pulmonary artery isolated from non-cystic fibrosis (PAEC) and CF human lungs (CF-PAEC), under static conditions or physiological shear. CFTR activity, clearly detected in HUVEC and PAEC, was markedly reduced in CF-PAEC. CFTR blockade increased endothelial permeability to macromolecules and reduced trans‑endothelial electrical resistance (TEER). Consistent with this, CF-PAEC displayed lower TEER compared to PAEC. Under shear, CFTR blockade reduced VE-cadherin and p120 catenin membrane expression and triggered the formation of paxillin- and vinculin-enriched membrane blebs that evolved in shrinking of the cell body and disruption of cell-cell contacts. These changes were accompanied by enhanced release of microvesicles, which displayed reduced capability to stimulate proliferation in recipient EC. CFTR blockade also suppressed insulin-induced NO generation by EC, likely by inhibiting eNOS and AKT phosphorylation, whereas it enhanced IL-8 release. Remarkably, phosphodiesterase inhibitors in combination with a β2 adrenergic receptor agonist corrected functional and morphological changes triggered by CFTR dysfunction in EC. Our results uncover regulatory functions of CFTR in EC, suggesting a physiological role of CFTR in the maintenance EC homeostasis and its involvement in pathogenetic aspects of CF. Moreover, our findings open avenues for novel pharmacology to control endothelial dysfunction and its consequences in CF. •CFTR-loss-of-function impairs endothelial monolayer integrity.•Changes in VE-cadherin and p120 catenin expression are involved.•CF endothelial cells release an increased number of dysfunctional microvesicles.•CFTR-loss-of-function impairs eNOS function.•cAMP-increasing drugs exert protective actions.