PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia

Shun Hay Pun; Karla M. O’Neill; Kevin S. Edgar; Eleanor K. Gill; Arya Moez; Hojjat Naderi-Meshkin; Sudhir B. Malla; Michelle B. Hookham; Mohammed Alsaggaf; Vinuthna Vani Madishetti; Bianca Botezatu; William King; Coy Brunssen; Henning Morawietz; Philip D. Dunne; Derek P. Brazil; Reinhold J. Medina; Chris J. Watson; David J. Grieve

doi:10.3390/cells12182220

PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia

Shun Hay Pun, Karla M. O’Neill, Kevin S. Edgar, Eleanor K. Gill, Arya Moez, Hojjat Naderi-Meshkin, Sudhir B. Malla, Michelle B. Hookham, Mohammed Alsaggaf, Vinuthna Vani Madishetti, Bianca Botezatu, William King, Coy Brunssen, Henning Morawietz, Philip D. Dunne, Derek P. Brazil, Reinhold J. Medina, Chris J. Watson, David J. Grieve^*

^*Corresponding author for this work

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Abstract

Ischaemic cardiovascular disease is associated with tissue hypoxia as a significant determinant of angiogenic dysfunction and adverse remodelling. While cord blood-derived endothelial colony-forming cells (CB-ECFCs) hold clear therapeutic potential due to their enhanced angiogenic and proliferative capacity, their impaired functionality within the disease microenvironment represents a major barrier to clinical translation. The aim of this study was to define the specific contribution of NOX4 NADPH oxidase, which we previously reported as a key CB-ECFC regulator, to hypoxia-induced dysfunction and its potential as a therapeutic target. CB-ECFCs exposed to experimental hypoxia demonstrated downregulation of NOX4-mediated reactive oxygen species (ROS) signalling linked with a reduced tube formation, which was partially restored by NOX4 plasmid overexpression. siRNA knockdown of placenta-specific 8 (PLAC8), identified by microarray analysis as an upstream regulator of NOX4 in hypoxic versus normoxic CB-ECFCs, enhanced tube formation, NOX4 expression and hydrogen peroxide generation, and induced several key transcription factors associated with downstream Nrf2 signalling. Taken together, these findings indicated that activation of the PLAC8–NOX4 signalling axis improved CB-ECFC angiogenic functions in experimental hypoxia, highlighting this pathway as a potential target for protecting therapeutic cells against the ischaemic cardiovascular disease microenvironment.

Original language	English
Article number	2220
Number of pages	19
Journal	Cells
Volume	12
Issue number	18
DOIs	https://doi.org/10.3390/cells12182220
Publication status	Published - 06 Sept 2023

Keywords

NOX4 NADPH oxidase
endothelial colony-forming cells
angiogenesis
placenta-specific 8
reactive oxygen species
hypoxia

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia
© 2023 The Authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Final published version, 4.44 MBLicence: CC BY

Interaction between NOX4 signalling in endothelial colony-forming cells and coronary microvascular function as a key determinant of cardiac remodelling
Pun, S. H. (Author), Watson, C. (Supervisor), Grieve, D. (Supervisor) & O'Neill, K. (Supervisor), Dec 2024
Student thesis: Doctoral Thesis › Doctor of Philosophy

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Pun, S. H., O’Neill, K. M., Edgar, K. S., Gill, E. K., Moez, A., Naderi-Meshkin, H., Malla, S. B., Hookham, M. B., Alsaggaf, M., Madishetti, V. V., Botezatu, B., King, W., Brunssen, C., Morawietz, H., Dunne, P. D., Brazil, D. P., Medina, R. J., Watson, C. J., & Grieve, D. J. (2023). PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia. Cells, 12(18), Article 2220. https://doi.org/10.3390/cells12182220

@article{9aeda40ce0f141bab608effdeb3caf8f,

title = "PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia",

abstract = "Ischaemic cardiovascular disease is associated with tissue hypoxia as a significant determinant of angiogenic dysfunction and adverse remodelling. While cord blood-derived endothelial colony-forming cells (CB-ECFCs) hold clear therapeutic potential due to their enhanced angiogenic and proliferative capacity, their impaired functionality within the disease microenvironment represents a major barrier to clinical translation. The aim of this study was to define the specific contribution of NOX4 NADPH oxidase, which we previously reported as a key CB-ECFC regulator, to hypoxia-induced dysfunction and its potential as a therapeutic target. CB-ECFCs exposed to experimental hypoxia demonstrated downregulation of NOX4-mediated reactive oxygen species (ROS) signalling linked with a reduced tube formation, which was partially restored by NOX4 plasmid overexpression. siRNA knockdown of placenta-specific 8 (PLAC8), identified by microarray analysis as an upstream regulator of NOX4 in hypoxic versus normoxic CB-ECFCs, enhanced tube formation, NOX4 expression and hydrogen peroxide generation, and induced several key transcription factors associated with downstream Nrf2 signalling. Taken together, these findings indicated that activation of the PLAC8–NOX4 signalling axis improved CB-ECFC angiogenic functions in experimental hypoxia, highlighting this pathway as a potential target for protecting therapeutic cells against the ischaemic cardiovascular disease microenvironment.",

keywords = "NOX4 NADPH oxidase, endothelial colony-forming cells, angiogenesis, placenta-specific 8, reactive oxygen species, hypoxia",

author = "Pun, {Shun Hay} and O{\textquoteright}Neill, {Karla M.} and Edgar, {Kevin S.} and Gill, {Eleanor K.} and Arya Moez and Hojjat Naderi-Meshkin and Malla, {Sudhir B.} and Hookham, {Michelle B.} and Mohammed Alsaggaf and Madishetti, {Vinuthna Vani} and Bianca Botezatu and William King and Coy Brunssen and Henning Morawietz and Dunne, {Philip D.} and Brazil, {Derek P.} and Medina, {Reinhold J.} and Watson, {Chris J.} and Grieve, {David J.}",

year = "2023",

month = sep,

day = "6",

doi = "10.3390/cells12182220",

language = "English",

volume = "12",

journal = "Cells",

issn = "2073-4409",

publisher = "MDPI",

number = "18",

}

Pun, SH, O’Neill, KM , Edgar, KS, Gill, EK, Moez, A, Naderi-Meshkin, H , Malla, SB, Hookham, MB, Alsaggaf, M , Madishetti, VV, Botezatu, B, King, W, Brunssen, C, Morawietz, H, Dunne, PD , Brazil, DP, Medina, RJ, Watson, CJ & Grieve, DJ 2023, 'PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia', Cells, vol. 12, no. 18, 2220. https://doi.org/10.3390/cells12182220

TY - JOUR

T1 - PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia

AU - Pun, Shun Hay

AU - O’Neill, Karla M.

AU - Edgar, Kevin S.

AU - Gill, Eleanor K.

AU - Moez, Arya

AU - Naderi-Meshkin, Hojjat

AU - Malla, Sudhir B.

AU - Hookham, Michelle B.

AU - Alsaggaf, Mohammed

AU - Madishetti, Vinuthna Vani

AU - Botezatu, Bianca

AU - King, William

AU - Brunssen, Coy

AU - Morawietz, Henning

AU - Dunne, Philip D.

AU - Brazil, Derek P.

AU - Medina, Reinhold J.

AU - Watson, Chris J.

AU - Grieve, David J.

PY - 2023/9/6

Y1 - 2023/9/6

N2 - Ischaemic cardiovascular disease is associated with tissue hypoxia as a significant determinant of angiogenic dysfunction and adverse remodelling. While cord blood-derived endothelial colony-forming cells (CB-ECFCs) hold clear therapeutic potential due to their enhanced angiogenic and proliferative capacity, their impaired functionality within the disease microenvironment represents a major barrier to clinical translation. The aim of this study was to define the specific contribution of NOX4 NADPH oxidase, which we previously reported as a key CB-ECFC regulator, to hypoxia-induced dysfunction and its potential as a therapeutic target. CB-ECFCs exposed to experimental hypoxia demonstrated downregulation of NOX4-mediated reactive oxygen species (ROS) signalling linked with a reduced tube formation, which was partially restored by NOX4 plasmid overexpression. siRNA knockdown of placenta-specific 8 (PLAC8), identified by microarray analysis as an upstream regulator of NOX4 in hypoxic versus normoxic CB-ECFCs, enhanced tube formation, NOX4 expression and hydrogen peroxide generation, and induced several key transcription factors associated with downstream Nrf2 signalling. Taken together, these findings indicated that activation of the PLAC8–NOX4 signalling axis improved CB-ECFC angiogenic functions in experimental hypoxia, highlighting this pathway as a potential target for protecting therapeutic cells against the ischaemic cardiovascular disease microenvironment.

AB - Ischaemic cardiovascular disease is associated with tissue hypoxia as a significant determinant of angiogenic dysfunction and adverse remodelling. While cord blood-derived endothelial colony-forming cells (CB-ECFCs) hold clear therapeutic potential due to their enhanced angiogenic and proliferative capacity, their impaired functionality within the disease microenvironment represents a major barrier to clinical translation. The aim of this study was to define the specific contribution of NOX4 NADPH oxidase, which we previously reported as a key CB-ECFC regulator, to hypoxia-induced dysfunction and its potential as a therapeutic target. CB-ECFCs exposed to experimental hypoxia demonstrated downregulation of NOX4-mediated reactive oxygen species (ROS) signalling linked with a reduced tube formation, which was partially restored by NOX4 plasmid overexpression. siRNA knockdown of placenta-specific 8 (PLAC8), identified by microarray analysis as an upstream regulator of NOX4 in hypoxic versus normoxic CB-ECFCs, enhanced tube formation, NOX4 expression and hydrogen peroxide generation, and induced several key transcription factors associated with downstream Nrf2 signalling. Taken together, these findings indicated that activation of the PLAC8–NOX4 signalling axis improved CB-ECFC angiogenic functions in experimental hypoxia, highlighting this pathway as a potential target for protecting therapeutic cells against the ischaemic cardiovascular disease microenvironment.

KW - NOX4 NADPH oxidase

KW - endothelial colony-forming cells

KW - angiogenesis

KW - placenta-specific 8

KW - reactive oxygen species

KW - hypoxia

U2 - 10.3390/cells12182220

DO - 10.3390/cells12182220

M3 - Article

SN - 2073-4409

VL - 12

JO - Cells

JF - Cells

IS - 18

M1 - 2220

ER -

PLAC8-mediated activation of NOX4 signalling restores angiogenic function of endothelial colony-forming cells in experimental hypoxia

Abstract

Keywords

UN SDGs

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Student theses

Interaction between NOX4 signalling in endothelial colony-forming cells and coronary microvascular function as a key determinant of cardiac remodelling

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