-
Tytuł:
-
Thromboxane A 2 receptor activation via G α13 -RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells.
-
Autorzy:
-
Eckenstaler R; Martin-Luther-University Halle-Wittenberg, Department of Clinical Pharmacy and Pharmacotherapy, Halle (Saale), Germany.
Ripperger A; Martin-Luther-University Halle-Wittenberg, Department of Clinical Pharmacy and Pharmacotherapy, Halle (Saale), Germany.
Hauke M; Martin-Luther-University Halle-Wittenberg, Department of Clinical Pharmacy and Pharmacotherapy, Halle (Saale), Germany.
Braun H; Martin-Luther-University Halle-Wittenberg, Department of Clinical Pharmacy and Pharmacotherapy, Halle (Saale), Germany.
Ergün S; Institute of Anatomy and Cell Biology, Julius-Maximilians-University, Würzburg, Germany.
Schwedhelm E; Institute of Clinical Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Benndorf RA; Martin-Luther-University Halle-Wittenberg, Department of Clinical Pharmacy and Pharmacotherapy, Halle (Saale), Germany. Electronic address: .
-
Źródło:
-
Biochemical pharmacology [Biochem Pharmacol] 2022 Jul; Vol. 201, pp. 115069. Date of Electronic Publication: 2022 May 04.
-
Typ publikacji:
-
Journal Article; Research Support, Non-U.S. Gov't
-
Język:
-
English
-
Imprint Name(s):
-
Publication: Oxford : Elsevier Science
Original Publication: Oxford, New York [etc.] Paragamon Press.
-
MeSH Terms:
-
GTP-Binding Protein alpha Subunits, G12-G13*/metabolism
Human Umbilical Vein Endothelial Cells*/cytology
Human Umbilical Vein Endothelial Cells*/metabolism
Lim Kinases*/metabolism
Receptors, Thromboxane A2, Prostaglandin H2*/metabolism
rhoA GTP-Binding Protein*/genetics
rhoA GTP-Binding Protein*/metabolism
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology ; Humans ; Neovascularization, Physiologic ; Signal Transduction ; Vascular Endothelial Growth Factor A/metabolism ; rho-Associated Kinases ; rhoC GTP-Binding Protein
-
Contributed Indexing:
-
Keywords: Angiogenic sprouting, cell tension, Thromboxane A(2) receptor; Endothelial cells; Endothelial dysfunction; G(α13); LIMK2; RhoA, RhoC, ROCK
-
Substance Nomenclature:
-
0 (Receptors, Thromboxane A2, Prostaglandin H2)
0 (VEGFA protein, human)
0 (Vascular Endothelial Growth Factor A)
124671-05-2 (RHOA protein, human)
76898-47-0 (15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid)
EC 2.7.11.1 (LIMK2 protein, human)
EC 2.7.11.1 (Lim Kinases)
EC 2.7.11.1 (rho-Associated Kinases)
EC 3.6.5.1 (GTP-Binding Protein alpha Subunits, G12-G13)
EC 3.6.5.2 (RHOC protein, human)
EC 3.6.5.2 (rhoA GTP-Binding Protein)
EC 3.6.5.2 (rhoC GTP-Binding Protein)
-
Entry Date(s):
-
Date Created: 20220507 Date Completed: 20220614 Latest Revision: 20220627
-
Update Code:
-
20240105
-
DOI:
-
10.1016/j.bcp.2022.115069
-
PMID:
-
35525325
-
We could previously show that thromboxane A 2 receptor (TP) activation inhibits the angiogenic capacity of human endothelial cells, but the underlying mechanisms remained unclear. Therefore, the aim of this study was to elucidate TP signal transduction pathways relevant to angiogenic sprouting of human endothelial cells. To clarify this matter, we used RNAi-mediated gene silencing as well as pharmacological inhibition of potential TP downstream targets in human umbilical vein endothelial cells (HUVEC) and VEGF-induced angiogenic sprouting of HUVEC spheroids in vitro as a functional read-out. In this experimental set-up, the TP agonist U-46619 completely blocked VEGF-induced angiogenic sprouting of HUVEC spheroids. Moreover, in live-cell analyses TP activation induced endothelial cell contraction, sprout retraction as well as endothelial cell tension and focal adhesion dysregulation of HUVEC. These effects were reversed by pharmacological TP inhibition or TP knockdown. Moreover, we identified a TP-G α13 -RhoA/C-ROCK-LIMK2-dependent signal transduction pathway to be relevant for U-46619-induced inhibition of VEGF-mediated HUVEC sprouting. In line with these results, U-46619-mediated TP activation potently induced RhoA and RhoC activity in live HUVEC as measured by FRET biosensors. Interestingly, pharmacological inhibition of ROCK and LIMK2 also normalized U-46619-induced endothelial cell tension and focal adhesion dysregulation of HUVEC. In summary, our work reveals mechanisms by which the TP may disturb angiogenic endothelial function in disease states associated with sustained endothelial TP activation.
(Copyright © 2022 Elsevier Inc. All rights reserved.)
