BACKGROUND/AIMS: Endothelial cell stiffness plays a key role in endothelium-dependent control of vascular tone and arterial blood pressure. Actin polymerization and distribution of microfilaments is essential for mechanical cell stiffness. Chorein, a protein encoded by the VPS13A gene, defective in chorea-acanthocytosis (ChAc), is involved in neuronal cell survival as well as cortical actin polymerization of erythrocytes and blood platelets. Chorein is expressed in a wide variety of further cells, yet nothing is known about the impact of chorein on cells other than neurons, erythrocytes and platelets. The present study explored whether chorein is expressed in human umbilical vein endothelial cells (HUVECs) and addressed the putative role of chorein in the regulation of cytoskeletal architecture, stiffness and survival of those cells. METHODS: In HUVECs with or without silencing of the VPS13A gene, VPS13A mRNA expression was determined utilizing quantitative RT-PCR, cytoskeletal organization visualized by confocal microscopy, G/F actin ratio and phosphorylation status of focal adhesion kinase quantified by western blotting, cell death determined by flow cytometry, mechanical properties studied by atomic force microscopy (AFM) and cell morphology analysed by scanning ion conductance microscopy (SICM). RESULTS: VPS13A mRNA expression was detectable in HUVECs. Silencing of the VPS13A gene attenuated the filamentous actin network, decreased the ratio of soluble G-actin over filamentous F-actin, reduced cell stiffness and changed cell morphology as compared to HUVECs silenced with negative control siRNA. These effects were paralleled by a significant decrease in FAK phosphorylation following VPS13A silencing. Moreover, silencing of the VPS13A gene increased caspase 3 activity and induced necrosis in HUVECs. CONCLUSIONS: Chorein is a novel regulator of cytoskeletal architecture, cell shape, mechanical stiffness and survival of vascular endothelial cells.
BACKGROUND/AIMS: Endothelial cell stiffness plays a key role in endothelium-dependent control of vascular tone and arterial blood pressure. Actin polymerization and distribution of microfilaments is essential for mechanical cell stiffness. Chorein, a protein encoded by the VPS13A gene, defective in chorea-acanthocytosis (ChAc), is involved in neuronal cell survival as well as cortical actin polymerization of erythrocytes and blood platelets. Chorein is expressed in a wide variety of further cells, yet nothing is known about the impact of chorein on cells other than neurons, erythrocytes and platelets. The present study explored whether chorein is expressed in human umbilical vein endothelial cells (HUVECs) and addressed the putative role of chorein in the regulation of cytoskeletal architecture, stiffness and survival of those cells. METHODS: In HUVECs with or without silencing of the VPS13A gene, VPS13A mRNA expression was determined utilizing quantitative RT-PCR, cytoskeletal organization visualized by confocal microscopy, G/F actin ratio and phosphorylation status of focal adhesion kinase quantified by western blotting, cell death determined by flow cytometry, mechanical properties studied by atomic force microscopy (AFM) and cell morphology analysed by scanning ion conductance microscopy (SICM). RESULTS:VPS13A mRNA expression was detectable in HUVECs. Silencing of the VPS13A gene attenuated the filamentous actin network, decreased the ratio of soluble G-actin over filamentous F-actin, reduced cell stiffness and changed cell morphology as compared to HUVECs silenced with negative control siRNA. These effects were paralleled by a significant decrease in FAK phosphorylation following VPS13A silencing. Moreover, silencing of the VPS13A gene increased caspase 3 activity and induced necrosis in HUVECs. CONCLUSIONS:Chorein is a novel regulator of cytoskeletal architecture, cell shape, mechanical stiffness and survival of vascular endothelial cells.
Authors: Wondwossen M Yeshaw; Marianne van der Zwaag; Francesco Pinto; Liza L Lahaye; Anita Ie Faber; Rubén Gómez-Sánchez; Amalia M Dolga; Conor Poland; Anthony P Monaco; Sven Cd van IJzendoorn; Nicola A Grzeschik; Antonio Velayos-Baeza; Ody Cm Sibon Journal: Elife Date: 2019-02-11 Impact factor: 8.140
Authors: Ana Mesquita; Elena Cardenal-Muñoz; Eunice Dominguez; Sandra Muñoz-Braceras; Beatriz Nuñez-Corcuera; Ben A Phillips; Luis C Tábara; Qiuhong Xiong; Roberto Coria; Ludwig Eichinger; Pierre Golstein; Jason S King; Thierry Soldati; Olivier Vincent; Ricardo Escalante Journal: Autophagy Date: 2016-10-07 Impact factor: 16.016
Authors: Zenobia B Mehta; Nicholas Fine; Timothy J Pullen; Matthew C Cane; Ming Hu; Pauline Chabosseau; Gargi Meur; Antonio Velayos-Baeza; Anthony P Monaco; Lorella Marselli; Piero Marchetti; Guy A Rutter Journal: Am J Physiol Endocrinol Metab Date: 2016-06-21 Impact factor: 4.310
Authors: Madhuri S Salker; Nicolas Schierbaum; Nour Alowayed; Yogesh Singh; Andreas F Mack; Christos Stournaras; Tilman E Schäffer; Florian Lang Journal: Sci Rep Date: 2016-07-11 Impact factor: 4.379