GPR68 Senses Flow and Is Essential for Vascular Physiology

Xu, Jie; Mathur, Jayanti; Vessières, Emilie; Hammack, Scott; Nonomura, Kazuteru; Favre, Julie; Grimaud, Linda; Petrus, Matt; Francisco, Allain; Li, Jingyuan; Lee, Van; Xiang, Fu li; Mainquist, James; Cahalan, Stuart M.; Orth, Anthony P.; Walker, John R.; Ma, Shang; Lukacs, Viktor; Bordone, Laura; Bandell, Michael; Laffitte, Bryan; Xu, Yan; Chien, Shu; Henrion, Didier; Patapoutian, Ardem

doi:10.1016/j.cell.2018.03.076

Cited by 233 publications

(244 citation statements)

References 61 publications

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“…The reason for this selection is because we found that dilation responses were relatively consistent across arteries of slightly different diameters and magnitudes of preconstriction at each flow rate (Fig. 1), and these response curves were similar to those found in many previous studies using stepped increases in flow in mouse mesenteric arteries (Loufrani et al 2002;Ohlmann et al 2005;Sennoun et al 2009;Priou et al 2010;Leonetti et al 2013;Tarjus et al 2017;Xu et al 2018), and more robust than found in some studies (Douglas et al 2008;Besnier et al 2018). Although the calculated shear stress was different between arteries at the start of flow (Initial Shear Stress), due to variation in diameters after preconstriction with phenylephrine, the final calculated shear stress (Final Shear Stress) values had much less variability as indicated by the smaller SEM ("Control" arteries in Tables 2 and 3).…”

Section: Limitationssupporting

confidence: 84%

“…1A in [Looft-Wilson et al 2008]) and the deep constriction allows observation of a larger magnitude flow-induced vasodilation response. Moreover, use of this agonist and this magnitude of preconstriction is typical when examining flow-induced dilation in these arteries (Loufrani et al 2002;Sennoun et al 2009;Tarjus et al 2017;Besnier et al 2018;Xu et al 2018). This phenylephrine dose results in eNOS phosphorylation changes (Looft-Wilson et al 2013), and it is possible that these phosphorylation changes are maximal.…”

Section: Limitationsmentioning

confidence: 99%

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Flow does not alter eNOS phosphoryation at Ser1179 or Thr495 in preconstricted mouse mesenteric arteries

et al. 2018

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In arteries, endothelium‐dependent vasodilatory agonists and flow‐induced shear stress cause vasodilation largely by activation of the endothelial enzyme eNOS, which generates nitric oxide that relaxes vascular smooth muscle. Agonists activate eNOS in part through increased phosphorylation at Ser1179 and decreased phosphorylation at Thr495. We previously found that preconstriction of intact, isolated mouse mesenteric arteries with phenylephrine also caused increased Ser1179 and decreased Thr495 eNOS phosphorylation, and sequential treatment with the vasodilatory agonist acetylcholine did not cause any further change in phosphorylation at these sites, despite producing vasodilation. The present study tests the hypothesis that luminal flow in these arteries preconstricted with phenylephrine also produces vasodilation without phosphorylation changes at these sites. First‐order mesenteric arteries, isolated from male C57/BL6 mice (7–20 weeks of age) anesthetized with pentobarbital (50 mg/kg, i.p.), were cannulated, pressurized, and treated with stepped increases in luminal flow (15–120 μL/min). Flow resulted in dilation that plateaued at ~60 μL/min (31.3 ± 3.0% dilation) and was significantly (P < 0.001) NOS‐dependent at all flow rates (determined by 10−4 mol/L L‐NAME treatment). In separate arteries, preconstriction with phenylephrine (10−5 mol/L) resulted in increased eNOS phosphorylation at Ser1179 (P < 0.05) and decreased phosphorylation at Thr495, but subsequent flow at 60 μL/min for 5 or 15 min did not cause further changes in phosphorylation, despite causing dilation. Thus, flow‐induced dilation does not require changes in these eNOS phosphorylation sites beyond those induced by alpha1‐adrenergic stimulation with phenylephrine, indicating that eNOS is activated by other mechanisms during acute flow‐induced dilation of preconstricted arteries.

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Section: Limitationssupporting

confidence: 84%

Section: Limitationsmentioning

confidence: 99%

Flow does not alter eNOS phosphoryation at Ser1179 or Thr495 in preconstricted mouse mesenteric arteries

et al. 2018

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“…We examined expression of GPCR along the aligned trajectories. The vasodilatory flow sensor gene Gpr68 59 first arises in pre-Art capillary EC and is retained in Art ( Fig. 7c ).…”

Section: Resultsmentioning

confidence: 99%

A molecular map of lymph node blood vascular endothelium at single cell resolution

Brulois

Rajaraman

Szade

et al. 2020

Preprint

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48 49 Blood vascular endothelial cells (BECs) control the immune response by regulating immune 50 cell recruitment, metabolite exchange and blood flow in lymphoid tissues. However, the 51 diversity of BEC and their origins during immune angiogenesis remain poorly understood. 52Here we profile transcriptomes of BEC from mouse peripheral lymph nodes and map key 53 phenotypes to the vasculature. Our analysis identifies multiple novel subsets including a 54 venous population whose gene signature predicts an unexpectedly selective role in myeloid 55 cell (vs lymphocyte) recruitment to the medulla, confirmed by 2 photon videomicroscopy. 56We define five phenotypes of capillary lining BEC including a capillary resident regenerative 57 population (CRP) that displays stem cell and migratory gene signatures and contributes to58 homeostatic BEC turnover and to vascular neogenesis after immunization. Trajectory 59 analyses reveal retention of developmental programs along a progression of cellular 60 phenotypes from CRP to mature venous and arterial BEC subsets. Overall, our single cell 61 atlas provides a molecular blueprint of the lymph node blood vasculature and defines subset 62 specialization for immune cell recruitment and vascular homeostasis. 63 64 Introduction 65The vascular endothelium lining blood vessels regulates exchange of oxygen and 66 metabolites between the blood vascular compartment and tissues. In lymph nodes (LN), 67 additionally, the endothelium plays essential roles in controlling immune cell access. The 68 organization of the vasculature in LN is well characterized: Arteries entering at the hilus lead to 69 capillary arcades in the LN cortex; capillaries link to 'high endothelial venules' (HEV), the post 70 capillary venules that recruit lymphocytes from the blood 1 ; and the vasculature exits the lymph 71 node at the hilus. Upon immune challenge, lymph nodes increase in volume up to 10-fold or more 72 within days, and the vascular endothelium expands roughly proportionally from local precursors 1,2 . 73Vessel expansion involves extensive proliferation of both capillary and high endothelial cells 74 (HEC) 3 , without contribution from blood borne progenitors 4 ; and an increase in vessel numbers 75 through intussusceptive (splitting) angiogenesis 5 . However, the nature and extent of endothelial 76 cell diversity within LN remains incompletely understood. 77Single cell RNA profiling is a transformative technology for the identification of cell 78 diversity and elucidation of developmental and physical relationships. Here we provide a survey 79 of blood vessel endothelial cells (BEC) from mouse peripheral lymph nodes (PLN). We confirm 80 known features of high endothelium 6,7 , identify novel endothelial subsets, uncover unexpected 81 diversity among capillary cells and demonstrate a distinctive role of medullary veins in selective 82 myeloid cell recruitment. We define gene signatures and transcription regulatory factors for these 83 subsets, and map key subsets to the vasculature. We also identify a p...

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“…In Xenopus, v-atpase regulates pH in a regional manner to affect craniofacial morphogenesis (Adams et al, 2006;Vandenberg et al, 2011) . Notably, recent studies of GPR68 have shown that the receptor senses flow and stretch (Wei et al, 2018;Xu et al, 2018) . However, responses to both stimuli are dependent on mildly acidic conditions.…”

Section: Discussionmentioning

confidence: 99%

Coupling Metastasis to pH-Sensing GPR68 Using a Novel Small Molecule Inhibitor

Williams

Neitzel

Karki

et al. 2019

Preprint

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Acidic milieu is a hallmark of glycolytic metabolism which occurs in cancerous cells. The effect of the acidic environment on cancer progression can be categorized into effects on tumor cell survival, tumor metastasis, inflammatory response, and blood vessels. Yet the underlying signaling and cell biological underpinnings of these phenomena are not well understood. Here, we describe, ogremorphen, a first-in-class inhibitor of proton-sensing GPCR GPR68. Discovered from a chemical genetic zebrafish screen, ogremorphen perturbed neural crest migration. Furthermore, ogremorphen was shown to inhibit migration in a number of human melanoma lines, which derive from the neural crest. Phenome-wide association study identified a natural variant that is aberrantly active and is associated with metastasis of common cancers. Our study suggests that GPR68 activity is associated with an increased ability for cancer cell migration and contributes to metastasis.

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GPR68 Senses Flow and Is Essential for Vascular Physiology

Cited by 233 publications

References 61 publications

Flow does not alter eNOS phosphoryation at Ser1179 or Thr495 in preconstricted mouse mesenteric arteries

Flow does not alter eNOS phosphoryation at Ser1179 or Thr495 in preconstricted mouse mesenteric arteries

A molecular map of lymph node blood vascular endothelium at single cell resolution

Coupling Metastasis to pH-Sensing GPR68 Using a Novel Small Molecule Inhibitor

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