Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation

NADPH oxidase 4 (NOX4) regulates endothelial inflammation by producing hydrogen peroxide (H2O2) and to a lesser extent O2•-. The ratio of NOX4-derived H2O2 and O2•- can be altered by coenzyme Q (CoQ) mimics. Therefore, we hypothesize that cytochrome b5 reductase 3 (CYB5R3), a CoQ reductase abundant...

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Autores: Yuan, Shuai, Hahn, Scott A., Miller, Megan P., Sanker, Subramaniam, Calderon, Michael J., Sullivan, Mara, Dosunmu-Ogunbi, Atinuke M., Fazzari, Marco, Li, Yao, Reynolds, Michael, Wood, Katherine C., Croix, Claudette M. St, Stolz, Donna, Cifuentes-Pagano, Eugenia, Navas, Plácido, Shiva, Sruti, Schopfer, Francisco J., Pagano, Patrick J., Straub, Adam C.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/260091
Acceso en línea:http://hdl.handle.net/10261/260091
Access Level:acceso abierto
Palabra clave:CYB5R3
NOX4
CoQ
ROS
Inflammation
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spelling Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammationYuan, ShuaiHahn, Scott A.Miller, Megan P.Sanker, SubramaniamCalderon, Michael J.Sullivan, MaraDosunmu-Ogunbi, Atinuke M.Fazzari, MarcoLi, YaoReynolds, MichaelWood, Katherine C.Croix, Claudette M. StStolz, DonnaCifuentes-Pagano, EugeniaNavas, PlácidoShiva, SrutiSchopfer, Francisco J.Pagano, Patrick J.Straub, Adam C.CYB5R3NOX4CoQROSInflammationNADPH oxidase 4 (NOX4) regulates endothelial inflammation by producing hydrogen peroxide (H2O2) and to a lesser extent O2•-. The ratio of NOX4-derived H2O2 and O2•- can be altered by coenzyme Q (CoQ) mimics. Therefore, we hypothesize that cytochrome b5 reductase 3 (CYB5R3), a CoQ reductase abundant in vascular endothelial cells, regulates inflammatory activation. To examine endothelial CYB5R3 in vivo, we created tamoxifen-inducible endothelium-specific Cyb5r3 knockout mice (R3 KO). Radiotelemetry measurements of systolic blood pressure showed systemic hypotension in lipopolysaccharides (LPS) challenged mice, which was exacerbated in R3 KO mice. Meanwhile, LPS treatment caused greater endothelial dysfunction in R3 KO mice, evaluated by acetylcholine-induced vasodilation in the isolated aorta, accompanied by elevated mRNA expression of vascular adhesion molecule 1 (Vcam-1). Similarly, in cultured human aortic endothelial cells (HAEC), LPS and tumor necrosis factor α (TNF-α) induced VCAM-1 protein expression was enhanced by Cyb5r3 siRNA, which was ablated by silencing the Nox4 gene simultaneously. Moreover, super-resolution confocal microscopy indicated mitochondrial co-localization of CYB5R3 and NOX4 in HAECs. APEX2-based electron microscopy and proximity biotinylation also demonstrated CYB5R3's localization on the mitochondrial outer membrane and its interaction with NOX4, which was further confirmed by the proximity ligation assay. Notably, Cyb5r3 knockdown HAECs showed less total H2O2 but more mitochondrial O2•-. Using inactive or non-membrane bound active CYB5R3, we found that CYB5R3 activity and membrane translocation are needed for optimal generation of H2O2 by NOX4. Lastly, cells lacking the CoQ synthesizing enzyme COQ6 showed decreased NOX4-derived H2O2, indicating a requirement for endogenous CoQ in NOX4 activity. In conclusion, CYB5R3 mitigates endothelial inflammatory activation by assisting in NOX4-dependent H2O2 generation via CoQ.This work was supported by National Institutes of Health (NIH) R01 awards [R01 HL 133864 (A.C.S), R01 HL 128304 (A.C.S), R01 HL 149825 (A.C.S), R01 HL 153532 (A.C.S), R01 GM 125944 (F.J.S.), R01 DK 112854 (F.J.S.), 1S10OD021540-01 (Center for Biologic Imaging, University of Pittsburgh), 1S10RR019003-01 (Simon Watkins (S.W.)), 1S10RR025488-01 (S.W.), 1S10RR016236-01 (S.W)]. American Heart Association (AHA) [Established Investigator Award 19EIA34770095 (A.C.S.)], Post-doctoral Fellowship 19POST34410028 (S.Y.)]. American Society of Hematology (ASH) Minority Hematology Graduate Award (A.M.D-O.). Junta de Andalucía grant BIO-177 (P.N.), the FEDER Funding Program from the European Union and Spanish Ministry of Science, Innovation and Universities grant RED2018-102576-T (P.N.).ElsevierNational Institutes of Health (US)University of PittsburghWatkins FoundationAmerican Heart AssociationAmerican Society of HematologyJunta de AndalucíaEuropean CommissionAgencia Estatal de Investigación (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2022202220212022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/260091reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement//RED2018-102576-T/http://dx.doi.org/10.1016/j.redox.2021.102166Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2600912026-05-22T06:33:51Z
dc.title.none.fl_str_mv Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
title Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
spellingShingle Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
Yuan, Shuai
CYB5R3
NOX4
CoQ
ROS
Inflammation
title_short Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
title_full Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
title_fullStr Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
title_full_unstemmed Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
title_sort Cooperation between CYB5R3 and NOX4 via coenzyme Q mitigates endothelial inflammation
dc.creator.none.fl_str_mv Yuan, Shuai
Hahn, Scott A.
Miller, Megan P.
Sanker, Subramaniam
Calderon, Michael J.
Sullivan, Mara
Dosunmu-Ogunbi, Atinuke M.
Fazzari, Marco
Li, Yao
Reynolds, Michael
Wood, Katherine C.
Croix, Claudette M. St
Stolz, Donna
Cifuentes-Pagano, Eugenia
Navas, Plácido
Shiva, Sruti
Schopfer, Francisco J.
Pagano, Patrick J.
Straub, Adam C.
author Yuan, Shuai
author_facet Yuan, Shuai
Hahn, Scott A.
Miller, Megan P.
Sanker, Subramaniam
Calderon, Michael J.
Sullivan, Mara
Dosunmu-Ogunbi, Atinuke M.
Fazzari, Marco
Li, Yao
Reynolds, Michael
Wood, Katherine C.
Croix, Claudette M. St
Stolz, Donna
Cifuentes-Pagano, Eugenia
Navas, Plácido
Shiva, Sruti
Schopfer, Francisco J.
Pagano, Patrick J.
Straub, Adam C.
author_role author
author2 Hahn, Scott A.
Miller, Megan P.
Sanker, Subramaniam
Calderon, Michael J.
Sullivan, Mara
Dosunmu-Ogunbi, Atinuke M.
Fazzari, Marco
Li, Yao
Reynolds, Michael
Wood, Katherine C.
Croix, Claudette M. St
Stolz, Donna
Cifuentes-Pagano, Eugenia
Navas, Plácido
Shiva, Sruti
Schopfer, Francisco J.
Pagano, Patrick J.
Straub, Adam C.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv National Institutes of Health (US)
University of Pittsburgh
Watkins Foundation
American Heart Association
American Society of Hematology
Junta de Andalucía
European Commission
Agencia Estatal de Investigación (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv CYB5R3
NOX4
CoQ
ROS
Inflammation
topic CYB5R3
NOX4
CoQ
ROS
Inflammation
description NADPH oxidase 4 (NOX4) regulates endothelial inflammation by producing hydrogen peroxide (H2O2) and to a lesser extent O2•-. The ratio of NOX4-derived H2O2 and O2•- can be altered by coenzyme Q (CoQ) mimics. Therefore, we hypothesize that cytochrome b5 reductase 3 (CYB5R3), a CoQ reductase abundant in vascular endothelial cells, regulates inflammatory activation. To examine endothelial CYB5R3 in vivo, we created tamoxifen-inducible endothelium-specific Cyb5r3 knockout mice (R3 KO). Radiotelemetry measurements of systolic blood pressure showed systemic hypotension in lipopolysaccharides (LPS) challenged mice, which was exacerbated in R3 KO mice. Meanwhile, LPS treatment caused greater endothelial dysfunction in R3 KO mice, evaluated by acetylcholine-induced vasodilation in the isolated aorta, accompanied by elevated mRNA expression of vascular adhesion molecule 1 (Vcam-1). Similarly, in cultured human aortic endothelial cells (HAEC), LPS and tumor necrosis factor α (TNF-α) induced VCAM-1 protein expression was enhanced by Cyb5r3 siRNA, which was ablated by silencing the Nox4 gene simultaneously. Moreover, super-resolution confocal microscopy indicated mitochondrial co-localization of CYB5R3 and NOX4 in HAECs. APEX2-based electron microscopy and proximity biotinylation also demonstrated CYB5R3's localization on the mitochondrial outer membrane and its interaction with NOX4, which was further confirmed by the proximity ligation assay. Notably, Cyb5r3 knockdown HAECs showed less total H2O2 but more mitochondrial O2•-. Using inactive or non-membrane bound active CYB5R3, we found that CYB5R3 activity and membrane translocation are needed for optimal generation of H2O2 by NOX4. Lastly, cells lacking the CoQ synthesizing enzyme COQ6 showed decreased NOX4-derived H2O2, indicating a requirement for endogenous CoQ in NOX4 activity. In conclusion, CYB5R3 mitigates endothelial inflammatory activation by assisting in NOX4-dependent H2O2 generation via CoQ.
publishDate 2021
dc.date.none.fl_str_mv 2021
2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/260091
url http://hdl.handle.net/10261/260091
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement//RED2018-102576-T/
http://dx.doi.org/10.1016/j.redox.2021.102166
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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