RT Journal Article
SR Electronic
T1 Macrophage-specific NF-κB activation dynamics can segregate inflammatory bowel disease patients
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 535096
DO 10.1101/535096
A1 Stamatia Papoutsopoulou
A1 Michael D. Burkitt
A1 François Bergey
A1 Hazel England
A1 Rachael Hough
A1 Lorraine Schmidt
A1 David G Spiller
A1 Michael HR White
A1 Pawel Paszek
A1 Dean A. Jackson
A1 Vitor A.P. Martins Dos Santos
A1 Gernot Sellge
A1 D. Mark Pritchard
A1 Barry J. Campbell
A1 Werner Müller
A1 Chris S. Probert
YR 2019
UL http://biorxiv.org/content/early/2019/01/30/535096.abstract
AB 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.