TY - JOUR
T1 - Depletion of Arg/Abl2 improves endothelial cell adhesion and prevents vascular leak during inflammation
AU - Amado-Azevedo, Joana
AU - van Stalborch, Anne-Marieke D.
AU - Valent, Erik T.
AU - Nawaz, Kalim
AU - van Bezu, Jan
AU - Eringa, Etto C.
AU - Hoevenaars, Femke P. M.
AU - de Cuyper, Iris M.
AU - Hordijk, Peter L.
AU - van Hinsbergh, Victor W. M.
AU - van Nieuw Amerongen, Geerten P.
AU - Aman, Jurjan
AU - Margadant, Coert
N1 - Funding Information: We thank Tony Koleske, Ken Yamada, Arnoud Sonnenberg, and Roderick Beijersbergen for their generous gifts of reagents. We gratefully acknowledge technical support from Mark Hoogenboezem, Simon Tol, and Erik Mul (Central Facility, Sanquin Research, Amsterdam), and Ben Morris (Robotics and Screening Center, Netherlands Cancer Institute, Amsterdam). Arnoud Sonnenberg is thanked for critical reading of the manuscript. Funding Information: This work was supported by grants from the Netherlands Heart Foundation to G.P. van Nieuw Amerongen (#2003T3201) and J. Aman (#2014T064), the ESICM/ECCRN Young Investigator Award 2011 to J. Aman, and the Netherlands Organization for Scientific Research (NWO) to C. Margadant (ZonMW Veni 016.146.160). Funding Information: We thank Tony Koleske, Ken Yamada, Arnoud Sonnenberg, and Roderick Beijersbergen for their generous gifts of reagents. We gratefully acknowledge technical support from Mark Hoogenboezem, Simon Tol, and Erik Mul (Central Facility, Sanquin Research, Amsterdam), and Ben Morris (Robotics and Screening Center, Netherlands Cancer Institute, Amsterdam). Arnoud Sonnenberg is thanked for critical reading of the manuscript. Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/8
Y1 - 2021/8
N2 - Endothelial barrier disruption and vascular leak importantly contribute to organ dysfunction and mortality during inflammatory conditions like sepsis and acute respiratory distress syndrome. We identified the kinase Arg/Abl2 as a mediator of endothelial barrier disruption, but the role of Arg in endothelial monolayer regulation and its relevance in vivo remain poorly understood. Here we show that depletion of Arg in endothelial cells results in the activation of both RhoA and Rac1, increased cell spreading and elongation, redistribution of integrin-dependent cell-matrix adhesions to the cell periphery, and improved adhesion to the extracellular matrix. We further show that Arg is activated in the endothelium during inflammation, both in murine lungs exposed to barrier-disruptive agents, and in pulmonary microvessels of septic patients. Importantly, Arg-depleted endothelial cells were less sensitive to barrier-disruptive agents. Despite the formation of F-actin stress fibers and myosin light chain phosphorylation, Arg depletion diminished adherens junction disruption and intercellular gap formation, by reducing the disassembly of cell-matrix adhesions and cell retraction. In vivo, genetic deletion of Arg diminished vascular leak in the skin and lungs, in the presence of a normal immune response. Together, our data indicate that Arg is a central and non-redundant regulator of endothelial barrier integrity, which contributes to cell retraction and gap formation by increasing the dynamics of adherens junctions and cell-matrix adhesions in a Rho GTPase-dependent fashion. Therapeutic inhibition of Arg may provide a suitable strategy for the treatment of a variety of clinical conditions characterized by vascular leak.
AB - Endothelial barrier disruption and vascular leak importantly contribute to organ dysfunction and mortality during inflammatory conditions like sepsis and acute respiratory distress syndrome. We identified the kinase Arg/Abl2 as a mediator of endothelial barrier disruption, but the role of Arg in endothelial monolayer regulation and its relevance in vivo remain poorly understood. Here we show that depletion of Arg in endothelial cells results in the activation of both RhoA and Rac1, increased cell spreading and elongation, redistribution of integrin-dependent cell-matrix adhesions to the cell periphery, and improved adhesion to the extracellular matrix. We further show that Arg is activated in the endothelium during inflammation, both in murine lungs exposed to barrier-disruptive agents, and in pulmonary microvessels of septic patients. Importantly, Arg-depleted endothelial cells were less sensitive to barrier-disruptive agents. Despite the formation of F-actin stress fibers and myosin light chain phosphorylation, Arg depletion diminished adherens junction disruption and intercellular gap formation, by reducing the disassembly of cell-matrix adhesions and cell retraction. In vivo, genetic deletion of Arg diminished vascular leak in the skin and lungs, in the presence of a normal immune response. Together, our data indicate that Arg is a central and non-redundant regulator of endothelial barrier integrity, which contributes to cell retraction and gap formation by increasing the dynamics of adherens junctions and cell-matrix adhesions in a Rho GTPase-dependent fashion. Therapeutic inhibition of Arg may provide a suitable strategy for the treatment of a variety of clinical conditions characterized by vascular leak.
KW - Arg/Abl2
KW - Endothelial barrier function
KW - Inflammation
KW - Integrins
KW - VE-cadherin
KW - Vascular leak
UR - http://www.scopus.com/inward/record.url?scp=85103357268&partnerID=8YFLogxK
U2 - https://doi.org/10.1007/s10456-021-09781-x
DO - https://doi.org/10.1007/s10456-021-09781-x
M3 - Article
C2 - 33770321
SN - 0969-6970
VL - 24
SP - 677
EP - 693
JO - Angiogenesis
JF - Angiogenesis
IS - 3
ER -