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Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells

Standard

Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells. / Eckenstaler, Robert; Ripperger, Anne; Hauke, Michael; Braun, Heike; Ergün, Süleyman; Schwedhelm, Edzard; Benndorf, Ralf A.

In: BIOCHEM PHARMACOL, Vol. 201, 115069, 07.2022.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Eckenstaler, R, Ripperger, A, Hauke, M, Braun, H, Ergün, S, Schwedhelm, E & Benndorf, RA 2022, 'Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells', BIOCHEM PHARMACOL, vol. 201, 115069. https://doi.org/10.1016/j.bcp.2022.115069

APA

Eckenstaler, R., Ripperger, A., Hauke, M., Braun, H., Ergün, S., Schwedhelm, E., & Benndorf, R. A. (2022). Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells. BIOCHEM PHARMACOL, 201, [115069]. https://doi.org/10.1016/j.bcp.2022.115069

Vancouver

Eckenstaler R, Ripperger A, Hauke M, Braun H, Ergün S, Schwedhelm E et al. Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells. BIOCHEM PHARMACOL. 2022 Jul;201. 115069. https://doi.org/10.1016/j.bcp.2022.115069

Bibtex

@article{f19b3bcc280c4c1bb16729eae89a8a14,
title = "Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells",
abstract = "We could previously show that thromboxane A2 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-Gα13-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.",
keywords = "15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology, GTP-Binding Protein alpha Subunits, G12-G13/metabolism, Human Umbilical Vein Endothelial Cells/cytology, Humans, Lim Kinases/metabolism, Neovascularization, Physiologic, Receptors, Thromboxane A2, Prostaglandin H2/metabolism, Signal Transduction, Vascular Endothelial Growth Factor A/metabolism, rho-Associated Kinases, rhoA GTP-Binding Protein/genetics, rhoC GTP-Binding Protein",
author = "Robert Eckenstaler and Anne Ripperger and Michael Hauke and Heike Braun and S{\"u}leyman Erg{\"u}n and Edzard Schwedhelm and Benndorf, {Ralf A}",
note = "Copyright {\textcopyright} 2022 Elsevier Inc. All rights reserved.",
year = "2022",
month = jul,
doi = "10.1016/j.bcp.2022.115069",
language = "English",
volume = "201",
journal = "BIOCHEM PHARMACOL",
issn = "0006-2952",
publisher = "Elsevier Inc.",

}

RIS

TY - JOUR

T1 - Thromboxane A2 receptor activation via Gα13-RhoA/C-ROCK-LIMK2-dependent signal transduction inhibits angiogenic sprouting of human endothelial cells

AU - Eckenstaler, Robert

AU - Ripperger, Anne

AU - Hauke, Michael

AU - Braun, Heike

AU - Ergün, Süleyman

AU - Schwedhelm, Edzard

AU - Benndorf, Ralf A

N1 - Copyright © 2022 Elsevier Inc. All rights reserved.

PY - 2022/7

Y1 - 2022/7

N2 - We could previously show that thromboxane A2 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-Gα13-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.

AB - We could previously show that thromboxane A2 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-Gα13-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.

KW - 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology

KW - GTP-Binding Protein alpha Subunits, G12-G13/metabolism

KW - Human Umbilical Vein Endothelial Cells/cytology

KW - Humans

KW - Lim Kinases/metabolism

KW - Neovascularization, Physiologic

KW - Receptors, Thromboxane A2, Prostaglandin H2/metabolism

KW - Signal Transduction

KW - Vascular Endothelial Growth Factor A/metabolism

KW - rho-Associated Kinases

KW - rhoA GTP-Binding Protein/genetics

KW - rhoC GTP-Binding Protein

U2 - 10.1016/j.bcp.2022.115069

DO - 10.1016/j.bcp.2022.115069

M3 - SCORING: Journal article

C2 - 35525325

VL - 201

JO - BIOCHEM PHARMACOL

JF - BIOCHEM PHARMACOL

SN - 0006-2952

M1 - 115069

ER -