3PO inhibits inflammatory NFκB and stress-activated kinase signaling in primary human endothelial cells independently of its target PFKFB3

Jonas Aakre Wik; Peter Lundbäck; Lars la Cour Poulsen; Guttorm Haraldsen; Bjørn Steen Skålhegg; Johanna Hol

doi:10.1371/journal.pone.0229395

3PO inhibits inflammatory NFκB and stress-activated kinase signaling in primary human endothelial cells independently of its target PFKFB3

PLoS One. 2020 Mar 4;15(3):e0229395. doi: 10.1371/journal.pone.0229395. eCollection 2020.

Authors

Jonas Aakre Wik^{1

2

3}, Peter Lundbäck^{2

3}, Lars la Cour Poulsen^{1

3}, Guttorm Haraldsen^{1

2

3}, Bjørn Steen Skålhegg⁴, Johanna Hol^{1

3}

Affiliations

¹ Department of Pathology, Oslo University Hospital-Rikshospitalet, Oslo, Norway.
² Department of Pathology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
³ K.G Jebsen Inflammation Research Centre, Faculty of Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁴ Department of Nutrition, Division of Molecular Nutrition, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.

Abstract

Inhibition of the key glycolytic activator 6-phosphofructokinase 2/fructose-2,6-bisphosphatase-3 (PFKFB3) by 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3PO) strongly attenuates pathological angiogenesis in cancer and inflammation. In addition to modulating endothelial proliferation and migration, 3PO also dampens proinflammatory activation of endothelial cells and experimental inflammation in vivo, suggesting a potential for 3PO in the treatment of chronic inflammation. The aim of our study was to explore if the anti-inflammatory action of 3PO in human endothelial cells was mediated by inhibition of PFKFB3 and glycolysis and assess if other means of PFKFB3 inhibition reduced inflammatory activation in a similar manner. We found that 3PO caused a rapid and transient reduction in IL-1β- and TNF-induced phosphorylation of both IKKα/β and JNK, thus inhibiting signaling through the NFκB and the stress-activated kinase pathways. However, in contrast to 3PO-treatment, neither shRNA-mediated silencing of PFKFB3 nor treatment with the alternative PFKFB3 inhibitor 7,8-dihydroxy-3-(4-hydroxy-phenyl)-chromen-4-one (YN1) prevented cytokine-induced NFκB signaling and upregulation of the adhesion molecules VCAM-1 and E-selectin, implying off target effects of 3PO. Collectively, our results suggest that the anti-inflammatory action of 3PO in human endothelial cells is not limited to inhibition of PFKFB3 and cellular glycolysis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Human Umbilical Vein Endothelial Cells / cytology*
Human Umbilical Vein Endothelial Cells / drug effects*
Human Umbilical Vein Endothelial Cells / metabolism
Humans
I-kappa B Kinase / metabolism
Inflammation / metabolism
Inflammation / pathology
Interleukin-1beta / pharmacology
JNK Mitogen-Activated Protein Kinases / metabolism
MAP Kinase Signaling System / drug effects*
Mitogen-Activated Protein Kinases / metabolism*
NF-kappa B / metabolism*
Phosphofructokinase-2 / metabolism*
Phosphorylation / drug effects
Pyridines / pharmacology*
Tumor Necrosis Factor-alpha / pharmacology

Substances

3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one
Interleukin-1beta
NF-kappa B
Pyridines
Tumor Necrosis Factor-alpha
PFKFB3 protein, human
Phosphofructokinase-2
I-kappa B Kinase
JNK Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinases

Grants and funding

The study was funded by grants from the South-Eastern Norway Regional Health Authorities (https://www.helse-sorost.no/, 2014032 and 2017013, awarded JH) and the National Association for Public Health, Norway (https://nasjonalforeningen.no, 2016, awarded JH). The funders had no role in study design, data Collection and analysis, decision to publish, or preparation of the manuscript.