Lang, MB;
Leung, KY;
Greene, NDE;
Malone, KM;
Saginc, G;
Randi, AM;
Kiprianos, A;
... Mason, JC; + view all
(2023)
The actions of methotrexate on endothelial cells are dependent on the shear stress-induced regulation of one carbon metabolism.
Frontiers in Immunology
, 14
, Article 1209490. 10.3389/fimmu.2023.1209490.
Preview |
Text
fimmu-14-1209490.pdf - Published Version Download (13MB) | Preview |
Abstract
Objectives: The disease-modifying anti-rheumatic drug methotrexate (MTX) is recognized to reduce cardiovascular risk in patients with systemic inflammatory diseases. However, the molecular basis for these cardioprotective effects remains incompletely understood. This study evaluated the actions of low-dose MTX on the vascular endothelium. Methods: Human endothelial cells (EC) were studied under in vitro conditions relevant to inflammatory arthritis. These included culture in a pro-inflammatory microenvironment and exposure to fluid shear stress (FSS) using a parallel plate model. Respectively treated cells were analyzed by RNA sequencing and quantitative real-time PCR for gene expression, by immunoblotting for protein expression, by phosphokinase activity arrays, by flow cytometry for cell cycle analyses and by mass spectrometry to assess folate metabolite levels. Results: In static conditions, MTX was efficiently taken up by EC and caused cell cycle arrest concurrent with modulation of cell signaling pathways. These responses were reversed by folinic acid (FA), suggesting that OCM is a predominant target of MTX. Under FSS, MTX did not affect cell proliferation or pro-inflammatory gene expression. Exposure to FSS downregulated endothelial one carbon metabolism (OCM) as evidenced by decreased expression of key OCM genes and metabolites. Conclusion: We found that FSS significantly downregulated OCM and thereby rendered EC less susceptible to the effects of MTX treatment. The impact of shear stress on OCM suggested that MTX does not directly modulate endothelial function. The cardioprotective actions of MTX likely reflect direct actions on inflammatory cells and indirect benefit on the vascular endothelium.
| Type: | Article |
|---|---|
| Title: | The actions of methotrexate on endothelial cells are dependent on the shear stress-induced regulation of one carbon metabolism |
| Location: | Switzerland |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.3389/fimmu.2023.1209490 |
| Publisher version: | https://doi.org/10.3389/fimmu.2023.1209490 |
| Language: | English |
| Additional information: | Copyright © 2023 Lang, Leung, Greene, Malone, Saginc, Randi, Kiprianos, Maughan, Pericleous and Mason. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
| Keywords: | cardiovascular disease, endothelial cells, methotrexate, one carbon metabolism, shear stress, Humans, Methotrexate, Endothelial Cells, Antirheumatic Agents, Folic Acid, Carbon |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Biology and Cancer Dept |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10174416 |
Archive Staff Only
 |
View Item |
