20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension

Victor Garcia, Ankit Gilani, Brian Shkolnik, Varunkumar Pandey, Frank Fan Zhang, Rambabu Dakarapu, Shyam K. Gandham, Rami R. Reddy, Joan P. Graves, Artiom Gruzdev, Darryl C. Zeldin, Jorge H. Capdevila, John R. Falck, Michal Laniado Schwartzman

Research output: Research - peer-reviewArticle

  • 3 Citations

Abstract

RATIONALE:: 20-Hydroxyeicosatetraenoic acid (20-HETE), one of the principle cytochrome P450 (CYP) eicosanoids, is a potent vasoactive lipid whose vascular effects include stimulation of smooth muscle contractility, migration and proliferation, as well as endothelial cell dysfunction and inflammation. Increased levels of 20-HETE in experimental animals and in humans are associated with hypertension, stroke, myocardial infarction and vascular diseases. OBJECTIVE:: To date, a receptor/binding site for 20-HETE has been implicated based on the use of specific agonists and antagonists. The present study was undertaken to identify a receptor to which 20-HETE binds and through which it activates a signaling cascade that culminates in many of the functional outcomes attributed to 20-HETE in vitro and in vivo. METHODS AND RESULTS:: Using crosslinking analogs, click chemistry, binding assays, and functional assays, we identified GPR75, currently an orphan G-protein coupled receptor (GPCR), as a specific target of 20-HETE. In cultured human endothelial cells, 20-HETE binding to GPR75 stimulated Gαq/11 protein dissociation and increased inositol phosphate (IP-1) accumulation as well as GPCR-kinase interacting protein-1 (GIT1)-GPR75 binding, which further facilitated the c-Src-mediated transactivation of endothelial EGFR. This results in downstream signaling pathways which induce angiotensin-converting enzyme (ACE) expression and endothelial dysfunction. Knockdown of GPR75 or GIT1 prevented 20-HETE-mediated endothelial growth factor receptor (EGFR) phosphorylation and ACE induction. In vascular smooth muscle cells, GPR75-20-HETE pairing is associated with Gαq/11- and GIT1-mediated protein kinase C (PKC)-stimulated phosphorylation of MaxiKλ, linking GPR75 activation to 20-HETE-mediated vasoconstriction. GPR75 knockdown in a mouse model of 20-HETE-dependent hypertension prevented blood pressure elevation and 20-HETE-mediated increases in ACE expression, endothelial dysfunction, smooth muscle contractility and vascular remodeling. CONCLUSIONS:: This is the first report to identify a GPCR target for an eicosanoid of this class. The discovery of 20-HETE-GPR75 pairing presented here provides the molecular basis for the signaling and pathophysiological functions mediated by 20-HETE in hypertension and cardiovascular diseases.

LanguageEnglish (US)
JournalCirculation Research
DOIs
StateAccepted/In press - Mar 23 2017

Fingerprint

G-Protein-Coupled Receptors
Blood Vessels
Hypertension
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Peptidyl-Dipeptidase A
Vascular Endothelial Growth Factor Receptor
Eicosanoids
Protein Kinases
Smooth Muscle
Endothelial Cells
Phosphorylation
G-Protein-Coupled Receptor Kinases
Receptor-Interacting Protein Serine-Threonine Kinases
Click Chemistry
Enzyme Induction
Vasoconstriction
Cardiomyopathies
Vascular Diseases
Vascular Smooth Muscle
Cytochrome P-450 Enzyme System

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Garcia, V., Gilani, A., Shkolnik, B., Pandey, V., Zhang, F. F., Dakarapu, R., ... Schwartzman, M. L. (2017). 20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension. Circulation Research. DOI: 10.1161/CIRCRESAHA.116.310525

20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension. / Garcia, Victor; Gilani, Ankit; Shkolnik, Brian; Pandey, Varunkumar; Zhang, Frank Fan; Dakarapu, Rambabu; Gandham, Shyam K.; Reddy, Rami R.; Graves, Joan P.; Gruzdev, Artiom; Zeldin, Darryl C.; Capdevila, Jorge H.; Falck, John R.; Schwartzman, Michal Laniado.

In: Circulation Research, 23.03.2017.

Research output: Research - peer-reviewArticle

Garcia, V, Gilani, A, Shkolnik, B, Pandey, V, Zhang, FF, Dakarapu, R, Gandham, SK, Reddy, RR, Graves, JP, Gruzdev, A, Zeldin, DC, Capdevila, JH, Falck, JR & Schwartzman, ML 2017, '20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension' Circulation Research. DOI: 10.1161/CIRCRESAHA.116.310525
Garcia, Victor ; Gilani, Ankit ; Shkolnik, Brian ; Pandey, Varunkumar ; Zhang, Frank Fan ; Dakarapu, Rambabu ; Gandham, Shyam K. ; Reddy, Rami R. ; Graves, Joan P. ; Gruzdev, Artiom ; Zeldin, Darryl C. ; Capdevila, Jorge H. ; Falck, John R. ; Schwartzman, Michal Laniado. / 20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension. In: Circulation Research. 2017
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abstract = "RATIONALE:: 20-Hydroxyeicosatetraenoic acid (20-HETE), one of the principle cytochrome P450 (CYP) eicosanoids, is a potent vasoactive lipid whose vascular effects include stimulation of smooth muscle contractility, migration and proliferation, as well as endothelial cell dysfunction and inflammation. Increased levels of 20-HETE in experimental animals and in humans are associated with hypertension, stroke, myocardial infarction and vascular diseases. OBJECTIVE:: To date, a receptor/binding site for 20-HETE has been implicated based on the use of specific agonists and antagonists. The present study was undertaken to identify a receptor to which 20-HETE binds and through which it activates a signaling cascade that culminates in many of the functional outcomes attributed to 20-HETE in vitro and in vivo. METHODS AND RESULTS:: Using crosslinking analogs, click chemistry, binding assays, and functional assays, we identified GPR75, currently an orphan G-protein coupled receptor (GPCR), as a specific target of 20-HETE. In cultured human endothelial cells, 20-HETE binding to GPR75 stimulated Gαq/11 protein dissociation and increased inositol phosphate (IP-1) accumulation as well as GPCR-kinase interacting protein-1 (GIT1)-GPR75 binding, which further facilitated the c-Src-mediated transactivation of endothelial EGFR. This results in downstream signaling pathways which induce angiotensin-converting enzyme (ACE) expression and endothelial dysfunction. Knockdown of GPR75 or GIT1 prevented 20-HETE-mediated endothelial growth factor receptor (EGFR) phosphorylation and ACE induction. In vascular smooth muscle cells, GPR75-20-HETE pairing is associated with Gαq/11- and GIT1-mediated protein kinase C (PKC)-stimulated phosphorylation of MaxiKλ, linking GPR75 activation to 20-HETE-mediated vasoconstriction. GPR75 knockdown in a mouse model of 20-HETE-dependent hypertension prevented blood pressure elevation and 20-HETE-mediated increases in ACE expression, endothelial dysfunction, smooth muscle contractility and vascular remodeling. CONCLUSIONS:: This is the first report to identify a GPCR target for an eicosanoid of this class. The discovery of 20-HETE-GPR75 pairing presented here provides the molecular basis for the signaling and pathophysiological functions mediated by 20-HETE in hypertension and cardiovascular diseases.",
author = "Victor Garcia and Ankit Gilani and Brian Shkolnik and Varunkumar Pandey and Zhang, {Frank Fan} and Rambabu Dakarapu and Gandham, {Shyam K.} and Reddy, {Rami R.} and Graves, {Joan P.} and Artiom Gruzdev and Zeldin, {Darryl C.} and Capdevila, {Jorge H.} and Falck, {John R.} and Schwartzman, {Michal Laniado}",
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T1 - 20- HETE Signals Through G Protein-Coupled Receptor GPR75 (Gq) to Affect Vascular Function and Trigger Hypertension

AU - Garcia,Victor

AU - Gilani,Ankit

AU - Shkolnik,Brian

AU - Pandey,Varunkumar

AU - Zhang,Frank Fan

AU - Dakarapu,Rambabu

AU - Gandham,Shyam K.

AU - Reddy,Rami R.

AU - Graves,Joan P.

AU - Gruzdev,Artiom

AU - Zeldin,Darryl C.

AU - Capdevila,Jorge H.

AU - Falck,John R.

AU - Schwartzman,Michal Laniado

PY - 2017/3/23

Y1 - 2017/3/23

N2 - RATIONALE:: 20-Hydroxyeicosatetraenoic acid (20-HETE), one of the principle cytochrome P450 (CYP) eicosanoids, is a potent vasoactive lipid whose vascular effects include stimulation of smooth muscle contractility, migration and proliferation, as well as endothelial cell dysfunction and inflammation. Increased levels of 20-HETE in experimental animals and in humans are associated with hypertension, stroke, myocardial infarction and vascular diseases. OBJECTIVE:: To date, a receptor/binding site for 20-HETE has been implicated based on the use of specific agonists and antagonists. The present study was undertaken to identify a receptor to which 20-HETE binds and through which it activates a signaling cascade that culminates in many of the functional outcomes attributed to 20-HETE in vitro and in vivo. METHODS AND RESULTS:: Using crosslinking analogs, click chemistry, binding assays, and functional assays, we identified GPR75, currently an orphan G-protein coupled receptor (GPCR), as a specific target of 20-HETE. In cultured human endothelial cells, 20-HETE binding to GPR75 stimulated Gαq/11 protein dissociation and increased inositol phosphate (IP-1) accumulation as well as GPCR-kinase interacting protein-1 (GIT1)-GPR75 binding, which further facilitated the c-Src-mediated transactivation of endothelial EGFR. This results in downstream signaling pathways which induce angiotensin-converting enzyme (ACE) expression and endothelial dysfunction. Knockdown of GPR75 or GIT1 prevented 20-HETE-mediated endothelial growth factor receptor (EGFR) phosphorylation and ACE induction. In vascular smooth muscle cells, GPR75-20-HETE pairing is associated with Gαq/11- and GIT1-mediated protein kinase C (PKC)-stimulated phosphorylation of MaxiKλ, linking GPR75 activation to 20-HETE-mediated vasoconstriction. GPR75 knockdown in a mouse model of 20-HETE-dependent hypertension prevented blood pressure elevation and 20-HETE-mediated increases in ACE expression, endothelial dysfunction, smooth muscle contractility and vascular remodeling. CONCLUSIONS:: This is the first report to identify a GPCR target for an eicosanoid of this class. The discovery of 20-HETE-GPR75 pairing presented here provides the molecular basis for the signaling and pathophysiological functions mediated by 20-HETE in hypertension and cardiovascular diseases.

AB - RATIONALE:: 20-Hydroxyeicosatetraenoic acid (20-HETE), one of the principle cytochrome P450 (CYP) eicosanoids, is a potent vasoactive lipid whose vascular effects include stimulation of smooth muscle contractility, migration and proliferation, as well as endothelial cell dysfunction and inflammation. Increased levels of 20-HETE in experimental animals and in humans are associated with hypertension, stroke, myocardial infarction and vascular diseases. OBJECTIVE:: To date, a receptor/binding site for 20-HETE has been implicated based on the use of specific agonists and antagonists. The present study was undertaken to identify a receptor to which 20-HETE binds and through which it activates a signaling cascade that culminates in many of the functional outcomes attributed to 20-HETE in vitro and in vivo. METHODS AND RESULTS:: Using crosslinking analogs, click chemistry, binding assays, and functional assays, we identified GPR75, currently an orphan G-protein coupled receptor (GPCR), as a specific target of 20-HETE. In cultured human endothelial cells, 20-HETE binding to GPR75 stimulated Gαq/11 protein dissociation and increased inositol phosphate (IP-1) accumulation as well as GPCR-kinase interacting protein-1 (GIT1)-GPR75 binding, which further facilitated the c-Src-mediated transactivation of endothelial EGFR. This results in downstream signaling pathways which induce angiotensin-converting enzyme (ACE) expression and endothelial dysfunction. Knockdown of GPR75 or GIT1 prevented 20-HETE-mediated endothelial growth factor receptor (EGFR) phosphorylation and ACE induction. In vascular smooth muscle cells, GPR75-20-HETE pairing is associated with Gαq/11- and GIT1-mediated protein kinase C (PKC)-stimulated phosphorylation of MaxiKλ, linking GPR75 activation to 20-HETE-mediated vasoconstriction. GPR75 knockdown in a mouse model of 20-HETE-dependent hypertension prevented blood pressure elevation and 20-HETE-mediated increases in ACE expression, endothelial dysfunction, smooth muscle contractility and vascular remodeling. CONCLUSIONS:: This is the first report to identify a GPCR target for an eicosanoid of this class. The discovery of 20-HETE-GPR75 pairing presented here provides the molecular basis for the signaling and pathophysiological functions mediated by 20-HETE in hypertension and cardiovascular diseases.

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SN - 0009-7330

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