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Rnd3 as a Novel Target to Ameliorate Microvascular Leakage

BACKGROUND: Microvascular leakage of plasma proteins is a hallmark of inflammation that leads to tissue dysfunction. There are no current therapeutic strategies to reduce microvascular permeability. The purpose of this study was to identify the role of Rnd3, an atypical Rho family GTPase, in the con...

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Autores principales: Breslin, Jerome W., Daines, Dayle A., Doggett, Travis M., Kurtz, Kristine H., Souza‐Smith, Flavia M., Zhang, Xun E., Wu, Mack H., Yuan, Sarah Y.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4859298/
https://www.ncbi.nlm.nih.gov/pubmed/27048969
http://dx.doi.org/10.1161/JAHA.116.003336
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author Breslin, Jerome W.
Daines, Dayle A.
Doggett, Travis M.
Kurtz, Kristine H.
Souza‐Smith, Flavia M.
Zhang, Xun E.
Wu, Mack H.
Yuan, Sarah Y.
author_facet Breslin, Jerome W.
Daines, Dayle A.
Doggett, Travis M.
Kurtz, Kristine H.
Souza‐Smith, Flavia M.
Zhang, Xun E.
Wu, Mack H.
Yuan, Sarah Y.
author_sort Breslin, Jerome W.
collection PubMed
description BACKGROUND: Microvascular leakage of plasma proteins is a hallmark of inflammation that leads to tissue dysfunction. There are no current therapeutic strategies to reduce microvascular permeability. The purpose of this study was to identify the role of Rnd3, an atypical Rho family GTPase, in the control of endothelial barrier integrity. The potential therapeutic benefit of Rnd3 protein delivery to ameliorate microvascular leakage was also investigated. METHODS AND RESULTS: Using immunofluorescence microscopy, Rnd3 was observed primarily in cytoplasmic areas around the nuclei of human umbilical vein endothelial cells. Permeability to fluorescein isothiocyanate–albumin and transendothelial electrical resistance of human umbilical vein endothelial cell monolayers served as indices of barrier function, and RhoA, Rac1, and Cdc42 activities were determined using G‐LISA assays. Overexpression of Rnd3 significantly reduced the magnitude of thrombin‐induced barrier dysfunction, and abolished thrombin‐induced Rac1 inactivation. Depleting Rnd3 expression with siRNA significantly extended the time course of thrombin‐induced barrier dysfunction and Rac1 inactivation. Time‐lapse microscopy of human umbilical vein endothelial cells expressing GFP‐actin showed that co‐expression of mCherry‐Rnd3 attenuated thrombin‐induced reductions in local lamellipodia that accompany endothelial barrier dysfunction. Lastly, a novel Rnd3 protein delivery method reduced microvascular leakage in a rat model of hemorrhagic shock and resuscitation, assessed by both intravital microscopic observation of extravasation of fluorescein isothiocyanate–albumin from the mesenteric microcirculation, and direct determination of solute permeability in intact isolated venules. CONCLUSIONS: The data suggest that Rnd3 can shift the balance of RhoA and Rac1 signaling in endothelial cells. In addition, our findings suggest the therapeutic, anti‐inflammatory potential of delivering Rnd3 to promote endothelial barrier recovery during inflammatory challenge.
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spelling pubmed-48592982016-05-20 Rnd3 as a Novel Target to Ameliorate Microvascular Leakage Breslin, Jerome W. Daines, Dayle A. Doggett, Travis M. Kurtz, Kristine H. Souza‐Smith, Flavia M. Zhang, Xun E. Wu, Mack H. Yuan, Sarah Y. J Am Heart Assoc Original Research BACKGROUND: Microvascular leakage of plasma proteins is a hallmark of inflammation that leads to tissue dysfunction. There are no current therapeutic strategies to reduce microvascular permeability. The purpose of this study was to identify the role of Rnd3, an atypical Rho family GTPase, in the control of endothelial barrier integrity. The potential therapeutic benefit of Rnd3 protein delivery to ameliorate microvascular leakage was also investigated. METHODS AND RESULTS: Using immunofluorescence microscopy, Rnd3 was observed primarily in cytoplasmic areas around the nuclei of human umbilical vein endothelial cells. Permeability to fluorescein isothiocyanate–albumin and transendothelial electrical resistance of human umbilical vein endothelial cell monolayers served as indices of barrier function, and RhoA, Rac1, and Cdc42 activities were determined using G‐LISA assays. Overexpression of Rnd3 significantly reduced the magnitude of thrombin‐induced barrier dysfunction, and abolished thrombin‐induced Rac1 inactivation. Depleting Rnd3 expression with siRNA significantly extended the time course of thrombin‐induced barrier dysfunction and Rac1 inactivation. Time‐lapse microscopy of human umbilical vein endothelial cells expressing GFP‐actin showed that co‐expression of mCherry‐Rnd3 attenuated thrombin‐induced reductions in local lamellipodia that accompany endothelial barrier dysfunction. Lastly, a novel Rnd3 protein delivery method reduced microvascular leakage in a rat model of hemorrhagic shock and resuscitation, assessed by both intravital microscopic observation of extravasation of fluorescein isothiocyanate–albumin from the mesenteric microcirculation, and direct determination of solute permeability in intact isolated venules. CONCLUSIONS: The data suggest that Rnd3 can shift the balance of RhoA and Rac1 signaling in endothelial cells. In addition, our findings suggest the therapeutic, anti‐inflammatory potential of delivering Rnd3 to promote endothelial barrier recovery during inflammatory challenge. John Wiley and Sons Inc. 2016-04-05 /pmc/articles/PMC4859298/ /pubmed/27048969 http://dx.doi.org/10.1161/JAHA.116.003336 Text en © 2016 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial (http://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Research
Breslin, Jerome W.
Daines, Dayle A.
Doggett, Travis M.
Kurtz, Kristine H.
Souza‐Smith, Flavia M.
Zhang, Xun E.
Wu, Mack H.
Yuan, Sarah Y.
Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title_full Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title_fullStr Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title_full_unstemmed Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title_short Rnd3 as a Novel Target to Ameliorate Microvascular Leakage
title_sort rnd3 as a novel target to ameliorate microvascular leakage
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4859298/
https://www.ncbi.nlm.nih.gov/pubmed/27048969
http://dx.doi.org/10.1161/JAHA.116.003336
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