Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke

Cardiovascular diseases (CVDs) are the leading cause of death globally, with ischemic stroke being a major concern. An ischemic stroke occurs approximately every three seconds. One key cause of ischemic stroke is atherosclerotic plaque, which can block blood vessels or rupture, leading to a thrombot...

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Autor: Marqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1
Tipo de recurso: tesis doctoral
Fecha de publicación:2024
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:Dadun. Depósito Académico Digital de la Universidad de Navarra
Idioma:inglés
OAI Identifier:oai:dadun.unav.edu:10171/123823
Acceso en línea:https://hdl.handle.net/10171/123823
Access Level:acceso embargado
Palabra clave:Cultivo celular
Fisiología cardiovascular
Arterioesclerosis y Arteroesclerosis
Trombosis
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spelling Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic strokeMarqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1Cultivo celularFisiología cardiovascularArterioesclerosis y ArteroesclerosisTrombosisCardiovascular diseases (CVDs) are the leading cause of death globally, with ischemic stroke being a major concern. An ischemic stroke occurs approximately every three seconds. One key cause of ischemic stroke is atherosclerotic plaque, which can block blood vessels or rupture, leading to a thrombotic response and forming a thrombus that further occludes vessels. Oxidative stress, caused by an imbalance between the production and elimination of reactive oxygen species (ROS), is a primary molecular mechanism in atherothrombotic stroke. NADPH oxidases (NOXs) are enzymes that produce ROS, playing both physiological and pathological roles. Among the NOX family, NOX5 is unique, being regulated by intracellular calcium levels and not requiring complementary subunits to produce ROS. Although less studied due to its absence in rodent genomes, NOX5 is implicated in processes contributing to CVDs, including endothelial dysfunction and immune cell adhesion. NOX5 expression is elevated in atherosclerotic plaques and is increased by smoking. However, the detailed role of NOX5 in atherothrombotic stroke remains underexplored. We hypothesized that NOX5 contributes to atherothrombotic stroke through endothelial dysfunction, affecting processes such as extracellular matrix (ECM) modifications, thrombosis, blood-brain barrier (BBB) impairment, immune infiltration, and stroke onset. Our study aimed to uncover new molecular mechanisms and pathological processes driven by endothelial NOX5 in atherothrombotic stroke. To investigate, we used various models to examine the effects of NOX5 overexpression in endothelial cells. In vitro, we employed human aortic endothelial cells (teloHAEC), human brain microvascular endothelial cells (hCMEC/D3), and human umbilical vein endothelial cells (HUVEC), using adenovirus and expression plasmids for NOX5 overexpression. In vivo, we used a knock-in mouse model expressing human NOX5 in endothelial cells upon tamoxifen induction, subjecting these mice to FeCl3-induced carotid thrombosis and ischemic stroke. Additionally, in silico analyses were conducted to study proteomic changes in hCMEC/D3 cells infected with NOX5 adenovirus and thrombi from the carotid thrombosis model. Our results demonstrated that NOX5 overexpression in endothelial cells influences multiple stages of atherothrombotic stroke. Firstly, chronic NOX5 overexpression induces MMP-10 secretion, linked to ECM changes, atherosclerotic plaque instability, and stroke severity. Secondly, NOX5 promotes thrombosis through a sex-dependent mechanism: in females, via PGE2 secretion leading to inflammation, and in males, by activating neutrophils. Thirdly, NOX5 overexpression in BBB endothelial cells causes dysfunction, increasing apoptosis, cell migration, mitochondrial dysfunction, and reducing cell proliferation. Finally, NOX5 enhances immune cell infiltration into the brain by disrupting tight junctions and increasing immune cell adhesion, exacerbated by ischemic stroke. In conclusion, endothelial NOX5 plays a significant role in the pathophysiology of atherothrombotic ischemic stroke, acting as a pathological contributor or as a response to vascular damage. This identifies NOX5 as a potential therapeutic target for preventing atherothrombotic ischemic stroke.Zalba-Goñi, G. (Guillermo)Dadun. Depósito Académico Digital Universidad de Navarra20262026-01-0120262026-01-0120242024-06-21doctoral thesishttp://purl.org/coar/resource_type/c_db06info:eu-repo/semantics/doctoralThesisapplication/pdfhttps://hdl.handle.net/10171/123823reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglésengembargoed accesshttp://purl.org/coar/access_right/c_f1cfinfo:eu-repo/semantics/embargoedAccessoai:dadun.unav.edu:10171/1238232026-06-21T12:47:57Z
dc.title.none.fl_str_mv Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
title Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
spellingShingle Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
Marqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1
Cultivo celular
Fisiología cardiovascular
Arterioesclerosis y Arteroesclerosis
Trombosis
title_short Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
title_full Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
title_fullStr Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
title_full_unstemmed Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
title_sort Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
dc.creator.none.fl_str_mv Marqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1
author Marqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1
author_facet Marqués-Cantero, J. (Javier)|||/items/19f43fc2-15a8-4581-b206-48e41dac93f1
author_role author
dc.contributor.none.fl_str_mv Zalba-Goñi, G. (Guillermo)
Dadun. Depósito Académico Digital Universidad de Navarra
dc.subject.none.fl_str_mv Cultivo celular
Fisiología cardiovascular
Arterioesclerosis y Arteroesclerosis
Trombosis
topic Cultivo celular
Fisiología cardiovascular
Arterioesclerosis y Arteroesclerosis
Trombosis
description Cardiovascular diseases (CVDs) are the leading cause of death globally, with ischemic stroke being a major concern. An ischemic stroke occurs approximately every three seconds. One key cause of ischemic stroke is atherosclerotic plaque, which can block blood vessels or rupture, leading to a thrombotic response and forming a thrombus that further occludes vessels. Oxidative stress, caused by an imbalance between the production and elimination of reactive oxygen species (ROS), is a primary molecular mechanism in atherothrombotic stroke. NADPH oxidases (NOXs) are enzymes that produce ROS, playing both physiological and pathological roles. Among the NOX family, NOX5 is unique, being regulated by intracellular calcium levels and not requiring complementary subunits to produce ROS. Although less studied due to its absence in rodent genomes, NOX5 is implicated in processes contributing to CVDs, including endothelial dysfunction and immune cell adhesion. NOX5 expression is elevated in atherosclerotic plaques and is increased by smoking. However, the detailed role of NOX5 in atherothrombotic stroke remains underexplored. We hypothesized that NOX5 contributes to atherothrombotic stroke through endothelial dysfunction, affecting processes such as extracellular matrix (ECM) modifications, thrombosis, blood-brain barrier (BBB) impairment, immune infiltration, and stroke onset. Our study aimed to uncover new molecular mechanisms and pathological processes driven by endothelial NOX5 in atherothrombotic stroke. To investigate, we used various models to examine the effects of NOX5 overexpression in endothelial cells. In vitro, we employed human aortic endothelial cells (teloHAEC), human brain microvascular endothelial cells (hCMEC/D3), and human umbilical vein endothelial cells (HUVEC), using adenovirus and expression plasmids for NOX5 overexpression. In vivo, we used a knock-in mouse model expressing human NOX5 in endothelial cells upon tamoxifen induction, subjecting these mice to FeCl3-induced carotid thrombosis and ischemic stroke. Additionally, in silico analyses were conducted to study proteomic changes in hCMEC/D3 cells infected with NOX5 adenovirus and thrombi from the carotid thrombosis model. Our results demonstrated that NOX5 overexpression in endothelial cells influences multiple stages of atherothrombotic stroke. Firstly, chronic NOX5 overexpression induces MMP-10 secretion, linked to ECM changes, atherosclerotic plaque instability, and stroke severity. Secondly, NOX5 promotes thrombosis through a sex-dependent mechanism: in females, via PGE2 secretion leading to inflammation, and in males, by activating neutrophils. Thirdly, NOX5 overexpression in BBB endothelial cells causes dysfunction, increasing apoptosis, cell migration, mitochondrial dysfunction, and reducing cell proliferation. Finally, NOX5 enhances immune cell infiltration into the brain by disrupting tight junctions and increasing immune cell adhesion, exacerbated by ischemic stroke. In conclusion, endothelial NOX5 plays a significant role in the pathophysiology of atherothrombotic ischemic stroke, acting as a pathological contributor or as a response to vascular damage. This identifies NOX5 as a potential therapeutic target for preventing atherothrombotic ischemic stroke.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024-06-21
2026
2026-01-01
2026
2026-01-01
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv https://hdl.handle.net/10171/123823
url https://hdl.handle.net/10171/123823
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv embargoed access
http://purl.org/coar/access_right/c_f1cf
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/embargoedAccess
rights_invalid_str_mv embargoed access
http://purl.org/coar/access_right/c_f1cf
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str Dadun. Depósito Académico Digital de la Universidad de Navarra
collection Dadun. Depósito Académico Digital de la Universidad de Navarra
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