title: RNA Sequencing Analysis Detection of a Novel Pathway of Endothelial Dysfunction in Pulmonary Arterial Hypertension.
abstract: BACKGROUND
  Pulmonary arterial hypertension is characterized by endothelial dysregulation, but global changes in gene expression have not been related to perturbations in function.
  
  
  OBJECTIVE
  RNA sequencing was used to discriminate changes in transcriptomes of endothelial cells cultured from lungs of patients with idiopathic pulmonary arterial hypertension versus control subjects and to assess the functional significance of major differentially expressed transcripts.
  
  
  METHODS
  The endothelial transcriptomes from the lungs of seven control subjects and six patients with idiopathic pulmonary arterial hypertension were analyzed. Differentially expressed genes were related to bone morphogenetic protein type 2 receptor (BMPR2) signaling. Those down-regulated were assessed for function in cultured cells and in a transgenic mouse.
  
  
  RESULTS
  Fold differences in 10 genes were significant (P < 0.05), four increased and six decreased in patients versus control subjects. No patient was mutant for BMPR2. However, knockdown of BMPR2 by siRNA in control pulmonary arterial endothelial cells recapitulated 6 of 10 patient-related gene changes, including decreased collagen IV (COL4A1, COL4A2) and ephrinA1 (EFNA1). Reduction of BMPR2-regulated transcripts was related to decreased β-catenin. Reducing COL4A1, COL4A2, and EFNA1 by siRNA inhibited pulmonary endothelial adhesion, migration, and tube formation. In mice null for the EFNA1 receptor, EphA2, versus control animals, vascular endothelial growth factor receptor blockade and hypoxia caused more severe pulmonary hypertension, judged by elevated right ventricular systolic pressure, right ventricular hypertrophy, and loss of small arteries.
  
  
  CONCLUSIONS
  The novel relationship between BMPR2 dysfunction and reduced expression of endothelial COL4 and EFNA1 may underlie vulnerability to injury in pulmonary arterial hypertension.
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Rhodes
authors.affiliation: 2 Cardiovascular Institute.|4 Department of Genetics, and.
authors.forename: Hogune
authors.initials: H
authors.lastname: Im
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Aiqin
authors.initials: A
authors.lastname: Cao
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Jan K
authors.initials: JK
authors.lastname: Hennigs
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Lingli
authors.initials: L
authors.lastname: Wang
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Silin
authors.initials: S
authors.lastname: Sa
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Pin-I
authors.initials: PI
authors.lastname: Chen
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Nils P
authors.initials: NP
authors.lastname: Nickel
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Miyagawa
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Hopper
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Nancy F
authors.initials: NF
authors.lastname: Tojais
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Caiyun G
authors.initials: CG
authors.lastname: Li
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gu
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|5 Department of Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Edda
authors.initials: E
authors.lastname: Spiekerkoetter
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Zhaoying
authors.initials: Z
authors.lastname: Xian
authors.affiliation: 2 Cardiovascular Institute.|4 Department of Genetics, and.
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.affiliation: 2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Mingming
authors.initials: M
authors.lastname: Zhao
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Mark
authors.initials: M
authors.lastname: Kaschwich
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|5 Department of Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Del Rosario
authors.affiliation: 2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Daniel
authors.initials: D
authors.lastname: Bernstein
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|5 Department of Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Roham T
authors.initials: RT
authors.lastname: Zamanian
authors.affiliation: 2 Cardiovascular Institute.|5 Department of Medicine, Stanford University School of Medicine, Stanford, California.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
authors.affiliation: 2 Cardiovascular Institute.|4 Department of Genetics, and.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Snyder
authors.affiliation: 1 Vera Moulton Wall Center for Pulmonary Vascular Diseases.|2 Cardiovascular Institute.|3 Department of Pediatrics.
authors.forename: Marlene
authors.initials: M
authors.lastname: Rabinovitch
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D052006:Bone Morphogenetic Protein Receptors, Type II / Q000235:genetics*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000503:physiopathology*; D065627:Familial Primary Pulmonary Hypertension / Q000235:genetics* / Q000503:physiopathology; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008875:Middle Aged; D017423:Sequence Analysis, RNA / Q000379:methods*; D015398:Signal Transduction / Q000235:genetics; D059467:Transcriptome / Q000235:genetics; D055815:Young Adult
chemical_list: C498218:BMPR2 protein, human; C498219:Bmpr2 protein, mouse; D052006:Bone Morphogenetic Protein Receptors, Type II
keywords: collagen IV; ephrin; pulmonary hypertension; transcriptome; vascular endothelium