Papoutsopoulou, Stamatia ORCID: 0000-0001-6665-8508, Burkitt, Michael ORCID: 0000-0002-5055-6408, Bergey, François, England, Hazel, Hough, Rachael, Schmidt, Lorraine, Spiller, David ORCID: 0000-0003-2502-6787, 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.
Frontiers in Immunology, 10.
535096-.
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. <h4>Significance statement</h4> 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 |
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Uncontrolled Keywords: | Crohn's Disease, Autoimmune Disease, Inflammatory Bowel Disease, Clinical Research, Digestive Diseases, 2.1 Biological and endogenous factors, 2 Aetiology, Oral and gastrointestinal |
Depositing User: | Symplectic Admin |
Date Deposited: | 13 Nov 2019 15:11 |
Last Modified: | 14 Mar 2024 19:46 |
DOI: | 10.1101/535096 |
Open Access URL: | https://www.frontiersin.org/articles/10.3389/fimmu... |
Related URLs: | |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3061789 |