Activated CD47 promotes pulmonary arterial hypertension through targeting caveolin-1.

AIMS Pulmonary arterial hypertension (PAH) is a progressive lung disease characterized by pulmonary vasoconstriction and vascular remodelling, leading to increased pulmonary vascular resistance and right heart failure. Loss of nitric oxide (NO) signalling and increased endothelial nitric oxide synthase (eNOS)-derived oxidative stress are central to the pathogenesis of PAH, yet the mechanisms involved remain incompletely determined. In this study, we investigated the role activated CD47 plays in promoting PAH. METHODS AND RESULTS We report high-level expression of thrombospondin-1 (TSP1) and CD47 in the lungs of human subjects with PAH and increased expression of TSP1 and activated CD47 in experimental models of PAH, a finding matched in hypoxic human and murine pulmonary endothelial cells. In pulmonary endothelial cells CD47 constitutively associates with caveolin-1 (Cav-1). Conversely, in hypoxic animals and cell cultures activation of CD47 by TSP1 disrupts this constitutive interaction, promoting eNOS-dependent superoxide production, oxidative stress, and PAH. Hypoxic TSP1 null mice developed less right ventricular pressure and hypertrophy and markedly less arteriole muscularization compared with wild-type animals. Further, therapeutic blockade of CD47 activation in hypoxic pulmonary artery endothelial cells upregulated Cav-1, increased Cav-1CD47 co-association, decreased eNOS-derived superoxide, and protected animals from developing PAH. CONCLUSION Activated CD47 is upregulated in experimental and human PAH and promotes disease by limiting Cav-1 inhibition of dysregulated eNOS.

[1]  OlivierFeron,et al.  Moderate Caveolin-1 Downregulation Prevents NADPH Oxidase–Dependent Endothelial Nitric Oxide Synthase Uncoupling by Angiotensin II in Endothelial Cells , 2011 .

[2]  D. Roberts,et al.  Thrombospondin-1 supports blood pressure by limiting eNOS activation and endothelial-dependent vasorelaxation. , 2010, Cardiovascular research.

[3]  R. Khanin,et al.  Dynamic Changes in Lung MicroRNA Profiles During the Development of Pulmonary Hypertension due to Chronic Hypoxia and Monocrotaline , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[4]  Rui Li,et al.  Gene delivery of cytochrome p450 epoxygenase ameliorates monocrotaline-induced pulmonary artery hypertension in rats. , 2010, American journal of respiratory cell and molecular biology.

[5]  D. Roberts,et al.  Thrombospondin-1/CD47 Blockade following Ischemia-Reperfusion Injury Is Tissue Protective , 2009, Plastic and reconstructive surgery.

[6]  Kurt R Stenmark,et al.  Animal models of pulmonary arterial hypertension: the hope for etiological discovery and pharmacological cure. , 2009, American journal of physiology. Lung cellular and molecular physiology.

[7]  M. Gladwin,et al.  Thrombospondin-1-CD47 blockade and exogenous nitrite enhance ischemic tissue survival, blood flow and angiogenesis via coupled NO-cGMP pathway activation. , 2009, Nitric oxide : biology and chemistry.

[8]  A. Malik,et al.  Persistent eNOS activation secondary to caveolin-1 deficiency induces pulmonary hypertension in mice and humans through PKG nitration. , 2009, The Journal of clinical investigation.

[9]  D. Mosher,et al.  Differential Interactions of Thrombospondin-1, -2, and -4 with CD47 and Effects on cGMP Signaling and Ischemic Injury Responses* , 2009, Journal of Biological Chemistry.

[10]  C. Schwencke,et al.  Chronic NOS inhibition prevents adverse lung remodeling and pulmonary arterial hypertension in caveolin-1 knockout mice. , 2008, Pulmonary pharmacology & therapeutics.

[11]  W. Frishman,et al.  Pyrrolidine dithiocarbamate restores endothelial cell membrane integrity and attenuates monocrotaline-induced pulmonary artery hypertension. , 2008, American journal of physiology. Lung cellular and molecular physiology.

[12]  M. Toborek,et al.  The role of caveolin-1 in PCB77-induced eNOS phosphorylation in human-derived endothelial cells. , 2007, American journal of physiology. Heart and circulatory physiology.

[13]  W. Sessa,et al.  Reexpression of caveolin-1 in endothelium rescues the vascular, cardiac, and pulmonary defects in global caveolin-1 knockout mice , 2007, The Journal of experimental medicine.

[14]  S. Black,et al.  The role of nitric oxide synthase-derived reactive oxygen species in the altered relaxation of pulmonary arteries from lambs with increased pulmonary blood flow. , 2007, American journal of physiology. Heart and circulatory physiology.

[15]  P. Insel,et al.  Increased smooth muscle cell expression of caveolin‐1 and caveolae contribute to the pathophysiology of idiopathic pulmonary arterial hypertension , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  M. J. Wagner,et al.  Right-ventricular failure is associated with increased mitochondrial complex II activity and production of reactive oxygen species. , 2007, Cardiovascular research.

[17]  D. Wink,et al.  Thrombospondin-1 limits ischemic tissue survival by inhibiting nitric oxide-mediated vascular smooth muscle relaxation. , 2007, Blood.

[18]  D. Wink,et al.  CD47 Is Necessary for Inhibition of Nitric Oxide-stimulated Vascular Cell Responses by Thrombospondin-1* , 2006, Journal of Biological Chemistry.

[19]  B. Y. Chin,et al.  Carbon monoxide reverses established pulmonary hypertension , 2006, The Journal of experimental medicine.

[20]  S. Archer,et al.  An Abnormal Mitochondrial–Hypoxia Inducible Factor-1&agr;–Kv Channel Pathway Disrupts Oxygen Sensing and Triggers Pulmonary Arterial Hypertension in Fawn Hooded Rats: Similarities to Human Pulmonary Arterial Hypertension , 2006, Circulation.

[21]  N. Chesler,et al.  The Mechanobiology of Pulmonary Vascular Remodeling in the Congenital Absence of eNOS , 2006, Biomechanics and modeling in mechanobiology.

[22]  N. Voelkel,et al.  Loss of caveolin and heme oxygenase expression in severe pulmonary hypertension. , 2006, Chest.

[23]  D. Wink,et al.  Thrombospondin-1 inhibits endothelial cell responses to nitric oxide in a cGMP-dependent manner. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[24]  K. Rockett,et al.  Pivotal Role for Endothelial Tetrahydrobiopterin in Pulmonary Hypertension , 2005, Circulation.

[25]  J. Lawler,et al.  Endogenous thrombospondin-1 is not necessary for proliferation but is permissive for vascular smooth muscle cell responses to platelet-derived growth factor. , 2005, Matrix biology : journal of the International Society for Matrix Biology.

[26]  W. Sessa,et al.  Endothelial-specific expression of caveolin-1 impairs microvascular permeability and angiogenesis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Y. Fung,et al.  Cellular and molecular mechanisms of pulmonary vascular remodeling: role in the development of pulmonary hypertension. , 2004, Microvascular research.

[28]  H. Schröder,et al.  Aspirin induces nitric oxide release from vascular endothelium: a novel mechanism of action , 2004, British journal of pharmacology.

[29]  M. Humbert,et al.  Cellular and molecular pathobiology of pulmonary arterial hypertension. , 2004, Journal of the American College of Cardiology.

[30]  W. Sessa eNOS at a glance , 2004, Journal of Cell Science.

[31]  W. Seeger,et al.  Clinical classification of pulmonary hypertension. , 2001, Clinics in chest medicine.

[32]  H. Farber,et al.  Pulmonary hypertension as a risk factor for death in patients with sickle cell disease. , 2004, The New England journal of medicine.

[33]  K. Pritchard,et al.  Decreased association of HSP90 impairs endothelial nitric oxide synthase in fetal lambs with persistent pulmonary hypertension. , 2003, American journal of physiology. Heart and circulatory physiology.

[34]  U. Förstermann,et al.  Regulation of endothelial-type NO synthase expression in pathophysiology and in response to drugs. , 2002, Nitric oxide : biology and chemistry.

[35]  J. Ross,et al.  Defects in caveolin-1 cause dilated cardiomyopathy and pulmonary hypertension in knockout mice , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[36]  David S. Park,et al.  Caveolin-1-deficient Mice Are Lean, Resistant to Diet-induced Obesity, and Show Hypertriglyceridemia with Adipocyte Abnormalities* , 2002, The Journal of Biological Chemistry.

[37]  W. Sessa,et al.  Post-translational control of endothelial nitric oxide synthase: why isn't calcium/calmodulin enough? , 2001, The Journal of pharmacology and experimental therapeutics.

[38]  W. Sessa,et al.  Distinction between signaling mechanisms in lipid rafts vs. caveolae , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[39]  M. Drab,et al.  Loss of Caveolae, Vascular Dysfunction, and Pulmonary Defects in Caveolin-1 Gene-Disrupted Mice , 2001, Science.

[40]  A. Zeiher,et al.  Phosphorylation of the endothelial nitric oxide synthase at Ser‐1177 is required for VEGF‐induced endothelial cell migration , 2000, FEBS letters.

[41]  R. Busse,et al.  Signal transduction of eNOS activation. , 1999, Cardiovascular research.

[42]  W. Sessa,et al.  Regulation of endothelium-derived nitric oxide production by the protein kinase Akt , 1999, Nature.

[43]  J. Richalet,et al.  Pulmonary hypertension in high-altitude chronic hypoxia: response to nifedipine. , 1998, The European respiratory journal.

[44]  R. Hynes,et al.  Thrombospondin-1 is required for normal murine pulmonary homeostasis and its absence causes pneumonia. , 1998, The Journal of clinical investigation.

[45]  H. Ju,et al.  Direct Interaction of Endothelial Nitric-oxide Synthase and Caveolin-1 Inhibits Synthase Activity* , 1997, The Journal of Biological Chemistry.

[46]  Richard G. W. Anderson,et al.  Acylation Targets Endothelial Nitric-oxide Synthase to Plasmalemmal Caveolae (*) , 1996, The Journal of Biological Chemistry.

[47]  E H Bergofsky,et al.  Survival in Patients with Primary Pulmonary Hypertension: Results from a National Prospective Registry , 1991 .

[48]  C. Pernot [Pulmonary arterial hypertension]. , 1958, Revue medicale de Nancy.