NF-κB2 signalling in enteroids modulates enterocyte responses to secreted factors from bone marrow-derived dendritic cells

• Published in: Cell death & disease Vol. 10; no. 12; pp. 896 - 14
• Main Authors: Jones, Lauren G., Vaida, Andra, Thompson, Louise M., Ikuomola, Felix I., Caamaño, Jorge H., Burkitt, Michael D., Miyajima, Fabio, Williams, Jonathan M., Campbell, Barry J., Pritchard, D. Mark, Duckworth, Carrie A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 26.11.2019; Springer Nature B.V
• Subjects: 13/100; 13/106; 13/2; 13/21; 13/51; 14/63; 631/532/2118/2437; 64/60; 692/308/1426; 692/420/256/1980; 692/420/2780; 692/699/1503/257; 96/1; Animals; Antibodies; Apoptosis; Apoptosis - drug effects; Biochemistry; Biomedical and Life Sciences; Bone marrow; Bone Marrow Cells - cytology; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Caspase-3; Cell Biology; Cell Culture; Cell Degranulation - drug effects; Cell Proliferation - drug effects; Colitis; Colon; Culture Media, Conditioned - pharmacology; Degranulation; Dendritic cells; Dendritic Cells - drug effects; Dendritic Cells - metabolism; Enterocytes - cytology; Enterocytes - drug effects; Enterocytes - metabolism; Epithelial cells; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Immunology; Inflammatory bowel diseases; Interferon-gamma - pharmacology; Intestine, Small - metabolism; Life Sciences; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Mice, Inbred C57BL; Mucosa; NF-kappa B p52 Subunit - metabolism; NF-κB protein; Organoids - metabolism; Paneth Cells - drug effects; Paneth Cells - metabolism; Permeability; Reproducibility of Results; Sepsis; Signal Transduction; Small intestine; Transcription; Tumor necrosis factor; Tumor Necrosis Factor-alpha - pharmacology; γ-Interferon
• ISSN: 2041-4889; 2041-4889
• DOI: 10.1038/s41419-019-2129-5

Alternative pathway NF-κB signalling regulates susceptibility towards developing inflammatory bowel disease (IBD), colitis-associated cancer and sepsis-associated intestinal epithelial cell apoptosis and shedding. However, the cell populations responsible for the perturbed alternative pathway NF-κB signalling in intestinal mucosal pathology remain unclear. In order to investigate the contribution of the epithelial compartment, we have tested whether NF-κB2 regulated transcription in intestinal epithelial cells controls the intestinal epithelial response to cytokines that are known to disrupt intestinal barrier permeability. Enteroids were generated from the proximal, middle and distal regions of small intestine (SI) from C57BL/6J wild-type mice and displayed region-specific morphology that was maintained during sub-culture. Enteroids treated with 100 ng/mL TNF were compared with corresponding regions of SI from C57BL/6J mice treated systemically with 0.33 mg/kg TNF for 1.5 h. TNF-induced apoptosis in all regions of the intestine in vitro and in vivo but resulted in Paneth cell degranulation only in proximal tissue-derived SI and enteroids. TNF also resulted in increased enteroid sphericity (quantified as circularity from two-dimensional bright field images). This response was dose and time-dependent and correlated with active caspase-3 immunopositivity. Proximal tissue-derived enteroids generated from Nfκb2 − /− mice showed a significantly blunted circularity response following the addition of TNF, IFNγ, lipopolysaccharide (LPS) activated C57BL/6J-derived bone marrow-derived dendritic cells (BMDC) and secreted factors from LPS-activated BMDCs. However, Nfκb1 −/− mouse-derived enteroids showed no significant changes in response to these stimuli. In conclusion, the selection of SI region is important when designing enteroid studies as region-specific identity and response to stimuli such as TNF are maintained in culture. Intestinal epithelial cells are at least partially responsible for regulating their own fate by modulating NF-κB2 signalling in response to stimuli known to be involved in multiple intestinal and systemic diseases. Future studies are warranted to investigate the therapeutic potential of intestinal epithelial NF-κB2 inhibition.