Macrophage-specific NF-κB activation dynamics can segregate inflammatory bowel disease patients



Papoutsopoulou, Stamatia ORCID: 0000-0001-6665-8508, Burkitt, Michael, Bergey, François, England, Hazel, Hough, Rachael, Schmidt, Lorraine, Spiller, David, White, Michael HR, Paszek, Pawel, Jackson, Dean
et al (show 6 more authors) (2019) Macrophage-specific NF-κB activation dynamics can segregate inflammatory bowel disease patients.

Access the full-text of this item by clicking on the Open Access link.

Abstract

Abstract The heterogeneous nature of inflammatory bowel disease (IBD) presents challenges, particularly when choosing therapy. Activation of the NF-κB transcription factor is a highly-regulated, dynamic event in IBD pathogenesis. We expressed the human NF-κB/p65 subunit in blood-derived macrophages, using lentivirus. Confocal imaging of p65 activation revealed that a higher proportion of macrophages from Crohn’s patients responded to lipid-A compared to controls. In contrast, cells from ulcerative colitis (UC) patients exhibited a shorter duration of p65 nuclear localisation compared to healthy controls and Crohn’s donors. Using a similar lentivirus approach, NF-κB-regulated luciferase was expressed in patient macrophages, isolated from frozen peripheral blood mononuclear cell samples. Following activation, samples could be segregated into three clusters based on the NF-κB-regulated luciferase response. The majority of UC samples appeared in hypo-responsive cluster 1, with Crohn’s patients representing the majority of hyper-responsive cluster 3. A positive correlation was seen between NF-κB-induced luciferase activity and cytokine levels released to medium from stimulated macrophages, but not in serum or biopsy. Analysis of macrophage cytokine responses and patient metadata revealed a strong correlation between Crohn’s patients who smoked and hyper-activation of p65. These in vitro dynamic assays of NF-κB activation in blood-derived macrophages segregate IBD patients into groups with different phenotypes and therefore may help determine response to therapy. Significance statement This manuscript describes two dynamic assays of NF-κB activation in blood-derived macrophages that can segregate IBD patients into groups with different phenotypes. For the first time we introduce the use of dynamic measurements of a transcription factor activation as a method to stratify patients and we are confident that our approach will lead in future to early patient stratification and prediction of treatment outcome.

Item Type: Article
Depositing User: Symplectic Admin
Date Deposited: 13 Nov 2019 15:11
Last Modified: 25 Feb 2020 10:19
DOI: 10.1101/535096
Open Access URL: https://www.frontiersin.org/articles/10.3389/fimmu...
URI: http://livrepository.liverpool.ac.uk/id/eprint/3061789