NOX4 is a major regulator of cord blood-derived endothelial colony-forming cells which promotes post-ischaemic revascularization
Year: 2019
Type: article
Abstract: AIMS: Cord blood-derived endothelial colony-forming cells (CB-ECFCs) are a defined progenitor population with established roles in vascular homeostasis and angiogenesis, which possess low immunogenicity and high potential for allogeneic therapy and are highly sensitive to regulation by reactive oxygen species (ROS). The aim of this study was to define the precise role of the major ROS-producing enzyme, NOX4 NADPH oxidase, in CB-ECFC vasoreparative function. METHODS AND RESULTS: In vitro CB-ECFC migration (scratch-wound assay) and tubulogenesis (tube length, branch number) was enhanced by phorbol 12-myristate 13-acetate (PMA)-induced superoxide in a NOX-dependent manner. CB-ECFCs highly-expressed NOX4, which was further induced by PMA, whilst NOX4 siRNA and plasmid overexpression reduced and potentiated in vitro function, respectively. Increased ROS generation in NOX4-overexpressing CB-ECFCs (DCF fluorescence, flow cytometry) was specifically reduced by superoxide dismutase, highlighting induction of ROS-specific signalling. Laser Doppler imaging of mouse ischaemic hindlimbs at 7 days indicated that NOX4-knockdown CB-ECFCs inhibited blood flow recovery, which was enhanced by NOX4-overexpressing CB-ECFCs. Tissue analysis at 14 days revealed consistent alterations in vascular density (lectin expression) and eNOS protein despite clearance of injected CB-ECFCs, suggesting NOX4-mediated modulation of host tissue. Indeed, proteome array analysis indicated that NOX4-knockdown CB-ECFCs largely suppressed tissue angiogenesis, whilst NOX4-overexpressing CB-ECFCs up-regulated a number of pro-angiogenic factors specifically-linked with eNOS signalling, in parallel with equivalent modulation of NOX-dependent ROS generation, suggesting that CB-ECFC NOX4 signalling may promote host vascular repair. CONCLUSION: Taken together, these findings indicate a key role for NOX4 in CB-ECFCs, thereby highlighting its potential as a target for enhancing their reparative function through therapeutic priming to support creation of a pro-reparative microenvironment and effective post-ischaemic revascularization.
Source: Cardiovascular Research
DOI: 10.1093/cvr/cvz090
Authors: Karla M O’Neill, David C Campbell, Kevin S Edgar, Eleanor K Gill, Arya Moez, Kiran J McLoughlin, Christina L O’Neill, Margaret Dellett, Ciarán J Hargey, Rawan A Abudalo, Michael O’Hare, Philip Doyle, Tinrui Toh, Joshua Khoo, June Wong, Cian M McCrudden, Marco Meloni, Coy Brunssen, Henning Morawietz, Mervin C Yoder, Denise M McDonald, Chris J Watson, Alan W Stitt, Andriana Margariti, Reinhold J Medina, David J Grieve
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,147
Awards: Endothelial NOX2 NADPH oxidase as a key driver of adverse cardiac remodelling associated with diabetes, Investigating the dynamic role of Quaking mRNA splicing in endothelial cell differentiation and its efficacy for vascular regeneration, Does Nox4 play a key role in direct reprogramming of fibroblasts into endothelial cells and underlie their ability to promote angiogenesis?, Influence of NOX4 NADPH oxidase on outgrowth endothelial cell function and their ability to promote angiogenesis in tissue ischaemia, Elucidating the mechanisms underpinning direct reprogramming of endothelial cells for use in regenerative medicine, The role of nitric oxide in vascular tip/stalk cell specification, G5113552736, Modulating glycolysis in vascular progenitors to improve tissue regeneration in ischaemic disease, Defining key mechanisms underlying NOX4 signalling in endothelial colony-forming cells towards improved vasoreparative capacity in ischaemic disease, Dedifferentiated or re-born again? Elucidating the chromatin remodelling mechanisms during endothelial cell reprogramming for cardiovascular therapy, Investigation of aberrant QKI splicing in diabetes and the specific role of QKI-7 as a novel target to reverse EC dysfunction
OpenAlex ID: W2931003500