title: Heterozygous null bone morphogenetic protein receptor type 2 mutations promote SRC kinase-dependent caveolar trafficking defects and endothelial dysfunction in pulmonary arterial hypertension.
abstract: Hereditary pulmonary arterial hypertension (HPAH) is a rare, fatal disease of the pulmonary vasculature. The majority of HPAH patients inherit mutations in the bone morphogenetic protein type 2 receptor gene (BMPR2), but how these promote pulmonary vascular disease is unclear. HPAH patients have features of pulmonary endothelial cell (PEC) dysfunction including increased vascular permeability and perivascular inflammation associated with decreased PEC barrier function. Recently, frameshift mutations in the caveolar structural protein gene Caveolin-1 (CAV-1) were identified in two patients with non-BMPR2-associated HPAH. Because caveolae regulate endothelial function and vascular permeability, we hypothesized that defects in caveolar function might be a common mechanism by which BMPR2 mutations promote pulmonary vascular disease. To explore this, we isolated PECs from mice carrying heterozygous null Bmpr2 mutations (Bmpr2(+/-)) similar to those found in the majority of HPAH patients. We show that Bmpr2(+/-) PECs have increased numbers and intracellular localization of caveolae and caveolar structural proteins CAV-1 and Cavin-1 and that these defects are reversed after blocking endocytosis with dynasore. SRC kinase is also constitutively activated in Bmpr2(+/-) PECs, and localization of CAV-1 to the plasma membrane is restored after treating Bmpr2(+/-) PECs with the SRC kinase inhibitor 3-(4-chlorophenyl)-1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP2). Late outgrowth endothelial progenitor cells isolated from HPAH patients show similar increased activation of SRC kinase. Moreover, Bmpr2(+/-) PECs have impaired endothelial barrier function, and barrier function is restored after treatment with PP2. These data suggest that heterozygous null BMPR2 mutations promote SRC-dependent caveolar trafficking defects in PECs and that this may contribute to pulmonary endothelial barrier dysfunction in HPAH patients.
authors.affiliation: From the Departments of Cell and Developmental Biology.
authors.forename: Allison R
authors.initials: AR
authors.lastname: Prewitt
authors.affiliation: Medicine.
authors.forename: Sampa
authors.initials: S
authors.lastname: Ghose
authors.affiliation: From the Departments of Cell and Developmental Biology.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Frump
authors.affiliation: Medicine.
authors.forename: Arumima
authors.initials: A
authors.lastname: Datta
authors.affiliation: Pediatrics, and.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Austin
authors.affiliation: Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee 37232.
authors.forename: Anne K
authors.initials: AK
authors.lastname: Kenworthy
authors.affiliation: Medicine, mark.de.caestecker@vanderbilt.edu.
authors.forename: Mark P
authors.initials: MP
authors.lastname: de Caestecker
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D052006:Bone Morphogenetic Protein Receptors, Type II / Q000235:genetics* / Q000378:metabolism; D021941:Caveolae / Q000378:metabolism / Q000473:pathology; D051242:Caveolin 1 / Q000235:genetics; D004705:Endocytosis / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D065627:Familial Primary Pulmonary Hypertension / Q000235:genetics* / Q000503:physiopathology; D016368:Frameshift Mutation; D006579:Heterozygote; D006801:Humans; D008168:Lung / Q000378:metabolism / Q000473:pathology; D051379:Mice; D021381:Protein Transport / Q000235:genetics*; D011743:Pyrimidines / Q000008:administration & dosage; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: C412373:AG 1879; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D051242:Caveolin 1; D011743:Pyrimidines; D019061:src-Family Kinases; C498218:BMPR2 protein, human; D052006:Bone Morphogenetic Protein Receptors, Type II
keywords: BMPR2; Caveolae; Caveolin; Endocytosis; Endothelial Dysfunction; Endothelial Permeability; Late Outgrowth Endothelial Progenitor Cells; Pulmonary Hypertension; SRC