Derivation and characterisation of endothelial cells from patients with chronic thromboembolic pulmonary hypertension

Abstract

Pulmonary endarterectomy (PEA) resected material offers a unique opportunity to develop an in vitro endothelial cell model of chronic thromboembolic pulmonary hypertension (CTEPH). We aimed to comprehensively analyze the endothelial function, molecular signature, and mitochondrial profile of CTEPH-derived endothelial cells to better understand the pathophysiological mechanisms of endothelial dysfunction behind CTEPH, and to identify potential novel targets for the prevention and treatment of the disease. Isolated cells from specimens obtained at PEA (CTEPH-EC), were characterized based on morphology, phenotype, and functional analyses (in vitro and in vivo tubule formation, proliferation, apoptosis, and migration). Mitochondrial content, morphology, and dynamics, as well as high-resolution respirometry and oxidative stress, were also studied. CTEPH-EC displayed a hyperproliferative phenotype with an increase expression of adhesion molecules and a decreased apoptosis, eNOS activity, migration capacity and reduced angiogenic capacity in vitro and in vivo compared to healthy endothelial cells. CTEPH-EC presented altered mitochondrial dynamics, increased mitochondrial respiration and an unbalanced production of reactive oxygen species and antioxidants. Our study is the foremost comprehensive investigation of CTEPH-EC. Modulation of redox, mitochondrial homeostasis and adhesion molecule overexpression arise as novel targets and biomarkers in CTEPH.

Metabolic profile in endothelial cells of chronic thromboembolic pulmonary hypertension and pulmonary arterial hypertension

“…Additionally, our sample size is limited, and these results should be verified in a bigger patient group. Future work will aim to expand in PAH-EC, our recent results in oxygen consumption and mitochondrial functionality in CTEPH-EC, where we found an increased mitochondrial respiration activity in all the respiratory chain complexes compared to HPAEC 37 . Additionally , we aim to study the metabolism of pulmonary artery smooth muscle cells (PASMCs) of both diseases.…”

Hyperactive mTORC1 in lung mesenchyme induces endothelial cell dysfunction and pulmonary vascular remodeling

“…tive potential but reduced angiogenic capacity is a true feature of ECs or whether this is secondary to limitations of in vitro assays where ECs are neither exposed to the paracrine signaling nor experiencing biomechanical forces from the pulmonary ECM. To determine the effects of support cells and biochemical forces on PH are hyperproliferative, with increased migration but reduced angiogenic capacity on standard angiogenesis assays (25,26). However, how these ECs are affected by other pulmonary cell types and paracrine ligand/receptor signaling remains unclear.…”

Oxidative Stress and Antioxidant Therapy in Pulmonary Hypertension

“…CTEPH-derived endothelial cells (CTEPH-EC) presented a significant increase in oxidative stress levels, specifically mtROS production, and reduced expression of SOD-2 compared to healthy human pulmonary artery ECs used as control cells [41]. Nukala et al also showed that CTEPH-EC exhibited an increase in intracellular ROS, advanced oxidation protein products and total protein carbonyl content (PCO), and a downregulation of GPX4 and GPX1 proteins, demonstrating dysregulation of the oxidative stress response and highlighting the involvement of oxidative stress in CTEPH [42].…”