IL-1β stimulates a novel axis within the NFκB pathway in endothelial cells regulated by IKKα and TAK-1

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Craig, Rachel and McIntosh, Kathryn and Ho, Ka Ho and McCulloch, Ashley and Riley, Christopher and Lawson, Christopher and MacKay, Simon P. and Paul, Andrew and Coats, Paul and Plevin, Robin (2025) IL-1β stimulates a novel axis within the NFκB pathway in endothelial cells regulated by IKKα and TAK-1. Biochemical Pharmacology, 232. 116736. ISSN 0006-2952 (https://doi.org/10.1016/j.bcp.2024.116736)

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Abstract

In this study we examined the activation of the non-canonical NFκB signalling pathway in endothelial cells. In HUVECs, LIGHT stimulated a delayed induction of serine 866/870 p100 phosphorylation linked to p52 NFκB formation. Surprisingly, the canonical ligand, IL-1β, stimulated a rapid phosphorylation or p100 which was not associated with p52 formation. Inhibition of IKKα activity, using DN-IKKα adenovirus, IKKα siRNA or a novel first-in-class selective IKKα inhibitor, SU1261, revealed IL-1β induced p100 phosphorylation to be dependent on IKKα. In contrast, IKKβ inhibition was found to be without effect. The NIK inhibitor, CW15337, did not affect IL-1β induced p100 phosphorylation however, both p100 and pIKKα/β phosphorylation was substantially reduced by inhibition of the upstream kinase TAK-1, suggesting phosphorylation of p100 is mediated by IKKα from within the canonical NEMO/IKKβ /IKKα complex. IL-1β also stimulated a rapid increase in nuclear translocation of p52, which was not affected by NIK inhibition, suggesting a source of p52 independent of p100 processing. Inhibition of TAK-1 abolished p52 and p65 nuclear translocation in response to IL-1β. SiRNA deletion or inhibition with dominant-negative virus of IKKα activity partially reduced p52 translocation, however pharmacological inhibition of IKKα was without effect. Inhibition of IKKβ abolished both p52 and p65 translocation. Taken together these results show that IL-1β stimulates a novel IKKα −dependent axis within the non-canonical NFκB pathway in endothelial cells which is NIK-independent and regulated by TAK-1. However, this pathway is not primarily responsible for the early nuclear translocation of p52, which is dependent on IKKβ. Elucidation of both these new pathways may be significant for NFκB biology within the endothelium.

ORCID iDs

Craig, Rachel, McIntosh, Kathryn ORCID logoORCID: https://orcid.org/0000-0003-0222-3585, Ho, Ka Ho ORCID logoORCID: https://orcid.org/0000-0003-0461-4379, McCulloch, Ashley, Riley, Christopher, Lawson, Christopher ORCID logoORCID: https://orcid.org/0000-0002-0729-182X, MacKay, Simon P. ORCID logoORCID: https://orcid.org/0000-0001-8000-6557, Paul, Andrew ORCID logoORCID: https://orcid.org/0000-0001-5775-2332, Coats, Paul ORCID logoORCID: https://orcid.org/0000-0002-6035-675X and Plevin, Robin ORCID logoORCID: https://orcid.org/0000-0002-7849-1220;
CORE (COnnecting REpositories)