title: Endothelial-like cells in chronic thromboembolic pulmonary hypertension: crosstalk with myofibroblast-like cells.
abstract: BACKGROUND
  Chronic thromboembolic pulmonary hypertension (CTEPH) is characterized by intravascular thrombus formation in the pulmonary arteries.Recently, it has been shown that a myofibroblast cell phenotype was predominant within endarterectomized tissues from CTEPH patients. Indeed, our recent study demonstrated the existence of not only myofibroblast-like cells (MFLCs), but also endothelial-like cells (ELCs). Under in vitro conditions, a few transitional cells (co-expressing both endothelial- and SM-cell markers) were observed in the ELC population. We hypothesized that MFLCs in the microenvironment created by the unresolved clot may promote the endothelial-mesenchymal transition and/or induce endothelial cell (EC) dysfunction.
  
  
  METHODS
  We isolated cells from these tissues and identified them as MFLCs and ELCs. In order to test whether the MFLCs provide the microenvironment which causes EC alterations, ECs were incubated in serum-free medium conditioned by MFLCs, or were grown in co-culture with the MFLCs.
  
  
  RESULTS
  Our experiments demonstrated that MFLCs promoted the commercially available ECs to transit to other mesenchymal phenotypes and/or induced EC dysfunction through inactivation of autophagy, disruption of the mitochondrial reticulum, alteration of the SOD-2 localization, and decreased ROS production. Indeed, ELCs included a few transitional cells, lost the ability to form autophagosomes, and had defective mitochondrial structure/function. Moreover, rapamycin reversed the phenotypic alterations and the gene expression changes in ECs co-cultured with MFLCs, thus suggesting that this agent had beneficial therapeutic effects on ECs in CTEPH tissues.
  
  
  CONCLUSIONS
  It is possible that the microenvironment created by the stabilized clot stimulates MFLCs to induce EC alterations.
authors.affiliation: Department of Respirology (B2), Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. sakaos@faculty.chiba-u.jp
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Sakao
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Hao
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Tanabe
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Kasahara
authors.forename: Katsushi
authors.initials: K
authors.lastname: Kurosu
authors.forename: Koichiro
authors.initials: K
authors.lastname: Tatsumi
mesh_terms: D001343:Autophagy; D002450:Cell Communication / Q000187:drug effects; D002478:Cells, Cultured; D002908:Chronic Disease; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D004691:Endarterectomy; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D006976:Hypertension, Pulmonary / Q000209:etiology* / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology; D058628:Myofibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011655:Pulmonary Embolism / Q000150:complications* / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D051785:Smad Proteins / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors; D000099156:Superoxide Dismutase 2
chemical_list: D017077:Culture Media, Conditioned; D008869:Microtubule-Associated Proteins; D017346:Protein Serine-Threonine Kinases; D017382:Reactive Oxygen Species; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta; D020123:Sirolimus; D051785:Smad Proteins; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C477012:MAP1LC3A protein, human