title: RCAN1.4 regulates VEGFR-2 internalisation, cell polarity and migration in human microvascular endothelial cells.
abstract: Regulator of calcineurin 1 (RCAN1) is an endogenous inhibitor of the calcineurin pathway in cells. It is expressed as two isoforms in vertebrates: RCAN1.1 is constitutively expressed in most tissues, whereas transcription of RCAN1.4 is induced by several stimuli that activate the calcineurin-NFAT pathway. RCAN1.4 is highly upregulated in response to VEGF in human endothelial cells in contrast to RCAN1.1 and is essential for efficient endothelial cell migration and tubular morphogenesis. Here, we show that RCAN1.4 has a role in the regulation of agonist-stimulated VEGFR-2 internalisation and establishment of endothelial cell polarity. siRNA-mediated gene silencing revealed that RCAN1 plays a vital role in regulating VEGF-mediated cytoskeletal reorganisation and directed cell migration and sprouting angiogenesis. Adenoviral-mediated overexpression of RCAN1.4 resulted in increased endothelial cell migration. Antisense-mediated morpholino silencing of the zebrafish RCAN1.4 orthologue revealed a disrupted vascular development further confirming a role for the RCAN1.4 isoform in regulating vascular endothelial cell physiology. Our data suggest that RCAN1.4 plays a novel role in regulating endothelial cell migration by establishing endothelial cell polarity in response to VEGF.
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.|King Abdullah International Medical Research Center (KAIMRC)-Eastern Region, King Saud bin Abdulaziz University for Health Sciences, Al Hasa, 31982, Saudi Arabia.
authors.forename: Ahmad F
authors.initials: AF
authors.lastname: Alghanem
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.
authors.forename: Emma L
authors.initials: EL
authors.lastname: Wilkinson
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.
authors.forename: Maxine S
authors.initials: MS
authors.lastname: Emmett
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Aljasir
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK.
authors.forename: Katherine
authors.initials: K
authors.lastname: Holmes
authors.affiliation: Departments of Internal Medicine and Molecular Biology, University of Texas, Southwestern Medical Centre, Dallas, TX, USA.
authors.forename: Beverley A
authors.initials: BA
authors.lastname: Rothermel
authors.affiliation: Institute of Cardiovascular Sciences, The Medical School, University of Birmingham, Birmingham, B15 2TT, UK.
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Simms
authors.affiliation: Institute of Cardiovascular Sciences, The Medical School, University of Birmingham, Birmingham, B15 2TT, UK.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Heath
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK. m.j.cross@liv.ac.uk.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Cross
mesh_terms: D000818:Animals; D002465:Cell Movement; D016764:Cell Polarity; D003599:Cytoskeleton / Q000378:metabolism; D004268:DNA-Binding Proteins; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004705:Endocytosis; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008024:Ligands; D055806:Microvessels / Q000166:cytology*; D008954:Models, Biological; D009124:Muscle Proteins / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology
chemical_list: D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D009124:Muscle Proteins; D020033:Protein Isoforms; C108612:RCAN1 protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; Endothelial; Migration; Polarisation; RCAN1; VEGFR-2