Caveolin1 Deficiency: A Type I Endothelial Interferonopathy Associated with Pulmonary Arterial Hypertension

CAV1 loss-of-function mutations have been associated with the development of pulmonary arterial hypertension (PAH). CAV1 is an integral component of endothelial caveolae, specialized lipid rafts that attach to the actin cytoskeleton and modulate receptor/signal transduction coupling. CAV1 loss in pulmonary artery endothelial cells produced a proliferative, hypermigratory cellular phenotype with a disrupted cytoskeletal architecture, mirroring known features of PAH pathobiology. Gene expression on Human Pulmonary Arterial Endothelial Cells (PAECs) transfected with non-targeting siRNA control pool or siRNA (ON-TARGET plus CAV1) in the presence or absence of TNF-alpha was evaluated Human Pulmonary Arterial Endothelial Cells (PAECs) (Lonza Walkersville, MD) were seeded on type I collagen coated plates and cultured in endothelial basal media-2 (EBM-2) supplemented with growth factors (EGM™-2 SingleQuot kit) and 2% FBS (Lonza). PAECs (Passage 1) were transfected with non-targeting siRNA control pool (ON-TARGET plus Non-targeting pool; Dharmacon) or 10 nM siRNA (ON-TARGET plus CAV1; Dharmacon, Lafayette, CO) with DharmaFECT1 transfecting reagent (Dharmacon) as described in the manufacture’s protocol for 48 h follow by 4 h stimulation with or without TNF-alpha 0.5 ng/ml (Sigma, St. Louis, MO). Five complete independent experiments were analyzed by microarray.