title: CFTR and sphingolipids mediate hypoxic pulmonary vasoconstriction.
abstract: Hypoxic pulmonary vasoconstriction (HPV) optimizes pulmonary ventilation-perfusion matching in regional hypoxia, but promotes pulmonary hypertension in global hypoxia. Ventilation-perfusion mismatch is a major cause of hypoxemia in cystic fibrosis. We hypothesized that cystic fibrosis transmembrane conductance regulator (CFTR) may be critical in HPV, potentially by modulating the response to sphingolipids as mediators of HPV. HPV and ventilation-perfusion mismatch were analyzed in isolated mouse lungs or in vivo. Ca(2+) mobilization and transient receptor potential canonical 6 (TRPC6) translocation were studied in human pulmonary (PASMCs) or coronary (CASMCs) artery smooth muscle cells. CFTR inhibition or deficiency diminished HPV and aggravated ventilation-perfusion mismatch. In PASMCs, hypoxia caused CFTR to interact with TRPC6, whereas CFTR inhibition attenuated hypoxia-induced TRPC6 translocation to caveolae and Ca(2+) mobilization. Ca(2+) mobilization by sphingosine-1-phosphate (S1P) was also attenuated by CFTR inhibition in PASMCs, but amplified in CASMCs. Inhibition of neutral sphingomyelinase (nSMase) blocked HPV, whereas exogenous nSMase caused TRPC6 translocation and vasoconstriction that were blocked by CFTR inhibition. nSMase- and hypoxia-induced vasoconstriction, yet not TRPC6 translocation, were blocked by inhibition or deficiency of sphingosine kinase 1 (SphK1) or antagonism of S1P receptors 2 and 4 (S1P2/4). S1P and nSMase had synergistic effects on pulmonary vasoconstriction that involved TRPC6, phospholipase C, and rho kinase. Our findings demonstrate a central role of CFTR and sphingolipids in HPV. Upon hypoxia, nSMase triggers TRPC6 translocation, which requires its interaction with CFTR. Concomitant SphK1-dependent formation of S1P and activation of S1P2/4 result in phospholipase C-mediated TRPC6 and rho kinase activation, which conjointly trigger vasoconstriction.
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Christoph
authors.initials: C
authors.lastname: Tabeling
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Hanpo
authors.initials: H
authors.lastname: Yu
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Liming
authors.initials: L
authors.lastname: Wang
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Hannes
authors.initials: H
authors.lastname: Ranke
authors.affiliation: The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada; Department of Anesthesia, University of Toronto, Toronto, ON M5G 1E2, Canada;
authors.forename: Neil M
authors.initials: NM
authors.lastname: Goldenberg
authors.affiliation: The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Diana
authors.initials: D
authors.lastname: Zabini
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Elena
authors.initials: E
authors.lastname: Noe
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Adrienn
authors.initials: A
authors.lastname: Krauszman
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Birgitt
authors.initials: B
authors.lastname: Gutbier
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada;
authors.forename: Jun
authors.initials: J
authors.lastname: Yin
authors.affiliation: Rudolf-Boehm-Institute of Pharmacology and Toxicology, University of Leipzig, 04107 Leipzig, Germany;
authors.forename: Michael
authors.initials: M
authors.lastname: Schaefer
authors.affiliation: Institute for Chemistry, Humboldt University, 12489 Berlin, Germany;
authors.forename: Christoph
authors.initials: C
authors.lastname: Arenz
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Andreas C
authors.initials: AC
authors.lastname: Hocke
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Norbert
authors.initials: N
authors.lastname: Suttorp
authors.affiliation: Genetics of Development and Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Proia
authors.affiliation: Departments of Infectious Diseases and Pulmonary Medicine and.
authors.forename: Martin
authors.initials: M
authors.lastname: Witzenrath
authors.affiliation: Physiology, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany; The Keenan Research Centre for Biomedical Science of St. Michael's, Toronto, ON M5B 1W8, Canada; Departments of Physiology and Surgery, University of Toronto, Toronto, ON M5G 1E2, Canada kueblerw@smh.ca.
authors.forename: Wolfgang M
authors.initials: WM
authors.lastname: Kuebler
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D002518:Ceramides / Q000737:chemistry; D003331:Coronary Vessels / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D006801:Humans; D006976:Hypertension, Pulmonary / Q000378:metabolism*; D000860:Hypoxia / Q000473:pathology; D008168:Lung / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018181:Mice, Inbred CFTR; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D021381:Protein Transport; D011651:Pulmonary Artery / Q000378:metabolism; D049349:Receptors, Lysosphingolipid / Q000378:metabolism; D015398:Signal Transduction; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism; D050052:TRPC Cation Channels / Q000378:metabolism; D000074287:TRPC6 Cation Channel; D010738:Type C Phospholipases / Q000378:metabolism; D014661:Vasoconstriction; D054460:rho-Associated Kinases / Q000378:metabolism; D000099157:Sphingosine Kinase
chemical_list: D002118:Calcium; D002518:Ceramides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D010100:Oxygen; D017853:Phosphotransferases (Alcohol Group Acceptor); D049349:Receptors, Lysosphingolipid; D013108:Sphingomyelin Phosphodiesterase; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; D010738:Type C Phospholipases; D054460:rho-Associated Kinases; D000099157:Sphingosine Kinase; C505032:CFTR protein, human; C117032:TRPC6 protein, human; C519633:SMPD2 protein, human
keywords: ceramide; neutral sphingomyelinase; pulmonary hypertension; transient receptor potential canonical 6
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