title: G-Protein-Coupled Receptor 35 Mediates Human Saphenous Vein Vascular Smooth Muscle Cell Migration and Endothelial Cell Proliferation.
abstract: Vascular smooth muscle cell (VSMC) migration and proliferation is central to neointima formation in vein graft failure following coronary artery bypass. However, there are currently no pharmacological interventions that prevent vein graft failure through intimal occlusion. It is hence a therapeutic target. Here, we investigated the contribution of GPR35 to human VSMC and endothelial cell (EC) migration, using a scratch-wound assay, and also the contribution to proliferation, using MTS and BrdU assays, in in vitro models using recently characterized human GPR35 ortholog-selective small-molecule agonists and antagonists. Real-time PCR studies showed GPR35 to be robustly expressed in human VSMCs and ECs. Stimulation of GPR35, with either the human-selective agonist pamoic acid or the reference agonist zaprinast, promoted VSMC migration in the scratch-wound assay. These effects were blocked by coincubation with either of the human GPR35-specific antagonists, CID-2745687 or ML-145. These GPR35-mediated effects were produced by inducing alterations in the actin cytoskeleton via the Rho A/Rho kinase signaling axis. Additionally, the agonist ligands stimulated a proliferative response in ECs. These studies highlight the potential that small molecules that stimulate or block GPR35 activity can modulate vascular proliferation and migration. These data propose GPR35 as a translational therapeutic target in vascular remodeling.
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, UK.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: McCallum
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Mackenzie
authors.forename: Nina
authors.initials: N
authors.lastname: Divorty
authors.forename: Carolyn
authors.initials: C
authors.lastname: Clarke
authors.forename: Christian
authors.initials: C
authors.lastname: Delles
authors.forename: Graeme
authors.initials: G
authors.lastname: Milligan
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Nicklin
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000636:Aminosalicylic Acids / Q000494:pharmacology; D002465:Cell Movement; D049109:Cell Proliferation; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D057809:HEK293 Cells; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D009284:Naphthols / Q000494:pharmacology; D011688:Purinones / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012501:Saphenous Vein / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction; D053778:Thiazolidines / Q000494:pharmacology; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D013997:Time Factors; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: C577640:2-hydroxy-4-(4-(5-(2-methyl-3-phenylprop-2-enylidene)-4-oxo-2-sulfanylidene-1,3-thiazolidin-3-yl)butanoylamino)benzoic acid; D000636:Aminosalicylic Acids; C111379:GPR35 protein, human; D006835:Hydrazones; D009284:Naphthols; D011688:Purinones; D043562:Receptors, G-Protein-Coupled; D053778:Thiazolidines; C577639:methyl 5-((tert-butylcarbamothioylhydrazinylidene)methyl)-1-(2,4-difluorophenyl)pyrazole-4-carboxylate; C066737:RHOA protein, human; C004368:pamoic acid; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; C011145:zaprinast; D013890:Thiourea