Endothelial-to-Mesenchymal Transition in Pulmonary Hypertension

Abstract

P ulmonary arterial hypertension (PAH) is a rare disorder, with a prevalence of 15 to 50 patients per million in the population. It is characterized by remodeling of the precapillary pulmonary arteries, with endothelial cell dysfunction contributing to endothelial and pulmonary artery smooth muscle cell proliferation. This remodeling increases pulmonary vascular resistance and pulmonary arterial pressure (mean pulmonary arterial pressure ≥25 mm Hg and a pulmonary capillary wedge Background-The vascular remodeling responsible for pulmonary arterial hypertension (PAH) involves predominantly the accumulation of α-smooth muscle actin-expressing mesenchymal-like cells in obstructive pulmonary vascular lesions. Endothelial-to-mesenchymal transition (EndoMT) may be a source of those α-smooth muscle actin-expressing cells. Methods and Results-In situ evidence of EndoMT in human PAH was obtained by using confocal microscopy of multiple fluorescent stainings at the arterial level, and by using transmission electron microscopy and correlative light and electron microscopy at the ultrastructural level. Findings were confirmed by in vitro analyses of human PAH and control cultured pulmonary artery endothelial cells. In addition, the mRNA and protein signature of EndoMT was recognized at the arterial and lung level by quantitative real-time polymerase chain reaction and Western blot analyses. We confirmed our human observations in established animal models of pulmonary hypertension (monocrotaline and SuHx). After establishing the first genetically modified rat model linked to BMPR2 mutations (BMPR2 Δ140Ex1/+ rats), we demonstrated that EndoMT is linked to alterations in signaling of BMPR2, a gene that is mutated in 70% of cases of familial PAH and in 10% to 40% of cases of idiopathic PAH. We identified molecular actors of this pathological transition, including twist overexpression and vimentin phosphorylation. We demonstrated that rapamycin partially reversed the protein expression patterns of EndoMT, improved experimental PAH, and decreased the migration of human pulmonary artery endothelial cells, providing the proof of concept that EndoMT is druggable. Conclusions-EndoMT is linked to alterations in BPMR2 signaling and is involved in the occlusive vas cular remodeling of PAH, findings that may have therapeutic implications.

Pulmonary Vascular and Ventricular Dysfunction in the Susceptible Patient (2015 Grover Conference Series)

“…In PAH, endothelial cell phenotype switching to myofibroblasts occurs in parallel to the histological rearrangement of PAECs in remodeled pulmonary arterioles and is linked to dysregulated bone morphogenetic protein receptor type 2 (BMPR-2) signaling, cell proliferation, and, presumably, fibrosis. 154 However, diethylaminoethylcellulose chromatography and other sensitive methods have confirmed detectable expression levels of (fibrillar) collagen III and the profibrotic protein connective tissue growth factor (CTGF) in PAECs under basal conditions. 155,156 Importantly, however, CTGF-collagen III signaling is increased in PAECs exposed to hypoxia for 24 hours, 157 which is a time point well in advance of the 3-7-day duration necessary to complete EndMT.…”

Potassium Channel Subfamily K Member 3 (KCNK3) Contributes to the Development of Pulmonary Arterial Hypertension

“…26 We have recently demonstrated high expression of vimentin and of phosphorylated vimentin (Ser55) during the development of PAH. 13 Ser55 on vimentin is phosphorylated in various types of cells only during the early mitotic phase by Cdk1. 27 Vimentin has been shown to interact with phosphorylated extracellular signal-regulated kinase (ERK) 1/2 and to protect it against dephosphorylation, by guest on May 7, 2018 http://circ.ahajournals.org/ Downloaded from extending its mitotic potential.…”

Translating Research into Improved Patient Care in Pulmonary Arterial Hypertension

“…More recently, the first models of pulmonary veno-occlusive disease induced by alkylating agents have been described (42,43). Using advances in transgenic technologies, mutant mice and rats have been generated to mimic features of the most common genetic cause of PAH, namely BMPR2 loss of function (44). Because the generation and characterization of these new genetic models are expensive and time-consuming, they should ideally be shared through a public repository.…”