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Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis

Aims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, e...

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Autores principales: Vara, Dina, Watt, Joanna M., Fortunato, Tiago M., Mellor, Harry, Burgess, Matthew, Wicks, Kate, Mace, Kimberly, Reeksting, Shaun, Lubben, Anneke, Wheeler-Jones, Caroline P.D., Pula, Giordano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5725637/
https://www.ncbi.nlm.nih.gov/pubmed/28793782
http://dx.doi.org/10.1089/ars.2016.6869
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author Vara, Dina
Watt, Joanna M.
Fortunato, Tiago M.
Mellor, Harry
Burgess, Matthew
Wicks, Kate
Mace, Kimberly
Reeksting, Shaun
Lubben, Anneke
Wheeler-Jones, Caroline P.D.
Pula, Giordano
author_facet Vara, Dina
Watt, Joanna M.
Fortunato, Tiago M.
Mellor, Harry
Burgess, Matthew
Wicks, Kate
Mace, Kimberly
Reeksting, Shaun
Lubben, Anneke
Wheeler-Jones, Caroline P.D.
Pula, Giordano
author_sort Vara, Dina
collection PubMed
description Aims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, electron paramagnetic resonance, pull-down of dRP-interacting proteins, followed by immunoblotting, gene silencing of different NADPH oxidases (NOXs), and their regulatory cosubunits by small interfering RNA (siRNA) transfection, and experiments with inhibitors of the sugar transporter glucose transporter 1 (GLUT1) were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NOX2 complex, but not NOX4. Increased reactive oxygen species generation in response to NOX2 activity leads to redox-dependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces vascular endothelial growth factor receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA, and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and NOX2-dependent stimulation of angiogenesis by dRP were confirmed in excisional wound and Matrigel plug vascularization assays in vivo using NOX2(−/−) mice. Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex. Conclusions: This study describes a novel molecular mechanism underlying the proangiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and upregulation of VEGFR2. Antioxid. Redox Signal. 28, 110–130.
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spelling pubmed-57256372018-01-10 Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis Vara, Dina Watt, Joanna M. Fortunato, Tiago M. Mellor, Harry Burgess, Matthew Wicks, Kate Mace, Kimberly Reeksting, Shaun Lubben, Anneke Wheeler-Jones, Caroline P.D. Pula, Giordano Antioxid Redox Signal Original Research Communications Aims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, electron paramagnetic resonance, pull-down of dRP-interacting proteins, followed by immunoblotting, gene silencing of different NADPH oxidases (NOXs), and their regulatory cosubunits by small interfering RNA (siRNA) transfection, and experiments with inhibitors of the sugar transporter glucose transporter 1 (GLUT1) were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NOX2 complex, but not NOX4. Increased reactive oxygen species generation in response to NOX2 activity leads to redox-dependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces vascular endothelial growth factor receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA, and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and NOX2-dependent stimulation of angiogenesis by dRP were confirmed in excisional wound and Matrigel plug vascularization assays in vivo using NOX2(−/−) mice. Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex. Conclusions: This study describes a novel molecular mechanism underlying the proangiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and upregulation of VEGFR2. Antioxid. Redox Signal. 28, 110–130. Mary Ann Liebert, Inc. 2018-01-10 2018-01-10 /pmc/articles/PMC5725637/ /pubmed/28793782 http://dx.doi.org/10.1089/ars.2016.6869 Text en © Dina Vara et al., 2017; Published by Mary Ann Liebert, Inc. This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research Communications
Vara, Dina
Watt, Joanna M.
Fortunato, Tiago M.
Mellor, Harry
Burgess, Matthew
Wicks, Kate
Mace, Kimberly
Reeksting, Shaun
Lubben, Anneke
Wheeler-Jones, Caroline P.D.
Pula, Giordano
Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title_full Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title_fullStr Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title_full_unstemmed Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title_short Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis
title_sort direct activation of nadph oxidase 2 by 2-deoxyribose-1-phosphate triggers nuclear factor kappa b-dependent angiogenesis
topic Original Research Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5725637/
https://www.ncbi.nlm.nih.gov/pubmed/28793782
http://dx.doi.org/10.1089/ars.2016.6869
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