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Tura-Ceide 2017 Abstract MITOEAGLE Barcelona

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COST Action MITOEAGLE

Endothelial cells as a model for metabolic shift in chronic thromboembolic pulmonary hypertension.

Link: MitoEAGLE

Tura-Ceide O, Matias D, Smolders V, Piccari L, Moren C, Guitart-Mampel M, Garrabou G, Garcia-Lucio J, Paul T, Blanco I, Peinado VI, Castella M, Barbera JA (2017)

Event: MitoEAGLE Barcelona 2017

COST Action MITOEAGLE

Chronic thromboembolic pulmonary hypertension (CTEPH) involves thrombotic obstruction of pulmonary arteries, intimal thickening and remodelling of small vessels. The treatment of choice for symptomatic CTEPH is pulmonary endarterectomy (PEA). The material obtained in PEA offers the opportunity to unravel pathophysiological mechanisms potentially underlying CTEPH. Cancer-like metabolic shift towards a glycolytic metabolism (Warburg effect) and down-regulation of mitochondrial glucose oxidation in endothelial cells might explain their hyperproliferative phenotype and vascular changes occurring in CTEPH. We aimed to develop an in vitro model of CTEPH using patient-derived cell lines to assess potential mitochondrial disturbances and alterations in the mitochondrial respiratory system.

Isolated cell lines (n=10) from specimens obtained at PEA were confirmed as being endothelial cells based on cobblestone morphology, endothelial phenotype (flow cytometry, qRT-PCR, immunofluorescence) and functional analysis (tubule formation, proliferation and migration). Mitochondrial studies were also performed: mitochondrial membrane potential (MMP) and apoptosis/necrosis by flow cytometry and mitochondrial dynamics (MD) by confocal microscopy. Finally, preliminary attempts on mitochondrial respiration were performed in permeabilized cells by high-resolution respirometry (Oroboros Oxygraph-2k), following either coupling control protocol or by stimulating with specific substrates for all the complexes of mitochondrial respiratory chain.

Isolated cells maintained cobblestone morphology and stained positive for endothelial markers. Hyperproliferative phenotype was observed when compared with control human pulmonary artery endothelial cell lines (HPAE): fold expansion (1.56±0.08 vs. 0.8±0.05; p<0.002). Functionally, reduced capacity to form tubule structures was found (96±21 vs. 150±44; p<0.03). CTEPH cells showed lower rates of depolarized, lower levels of necrosis/apoptosis and MD showed pathological trends with respect to controls. Moreover, mitochondrial oxygen consumption in CTEPH cells showed differential pattern compared to controls.

Endothelial cells obtained from PEA material showed a hyperproliferative phenotype, impaired function and mitochondrial derangements that may play a role in the development of CTEPH after pulmonary embolism.


Bioblast editor: Kandolf G O2k-Network Lab: ES Barcelona Moren C


Labels: MiParea: Respiration 


Tissue;cell: Endothelial;epithelial;mesothelial cell  Preparation: Permeabilized cells 



HRR: Oxygraph-2k  Event: B3 


Affiliations

Tura-Ceide O(1,2), Matias D(1), Smolders V(1,5), Piccari L(1), Morén C(3), Guitart-Mampel M(3), Garrabou G(3), García-Lucio J(1), Paul T, Blanco I(1,2), Peinado VI(1,2), Castellà M(4), Barberà JA(1,2)
  1. Servei Pneumologia, Hospital Clínic-IDIBAPS, Univ Barcelona
  2. Centro Investigación Biomédica Red Enfermedades Respiratorias
  3. Muscle Research Mitochondrial Function Lab, Cellex-Inst d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Centro de Investigación Biomédica Red Enfermedades Raras (CIBERER)
  4. Dept Cardiovascular Surgery, Inst Clínic del Tòrax, Hospital Clínic, Univ Barcelona
  5. Dept Biochem Molecular Biol, Fac Biol, Univ Barcelona, Barcelona; Spain.





Support

Supported by BIOTRACK EU-IDIBAPS postdoctoral programme (OTC); PFIS predoctoral programme from ISCIII (JGL); SEPAR (OTC), SOCAP (OTC), FIS: PI15/00582 (JAB), CIBERER (CM, MGM, GG) and HORIZON 2020 – Marie Skłodowska-Curie grant – ITN-European Joint Doctorate (VS).