1. TNFα Induces LGR5+ Stem Cell Dysfunction In Patients With Crohn’s Disease
- Author
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Chnasu Lee, Woong-Yang Park, Sung Noh Hong, Minae An, Je-Gun Joung, Dong Kyung Chang, and Young-Ho Kim
- Subjects
Tumor Necrosis Factor-Alpha ,Prostaglandin E2 ,Cellular differentiation ,Necroptosis ,Population ,PBS, phosphate-buffered saline ,RC799-869 ,Biology ,Receptors, G-Protein-Coupled ,ER, endoplasmic reticulum ,Intestinal Stem Cell ,MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ,Crohn Disease ,DEG, differentially expressed gene ,CD, Crohn’s disease ,PGE2, prostaglandin E2 ,Humans ,Viability assay ,TUNEL, deoxyuride-5′-triphosphate biotin nick end labeling ,Intestinal Mucosa ,education ,ANOVA, analysis of variance ,NF-κB, nuclear factor kappa B ,Original Research ,scRNA-seq, single-cell RNA sequencing ,TNF, tumor necrosis factor ,education.field_of_study ,Hepatology ,CC3, cleaved caspase-3 ,Stem Cells ,Intestinal Organoid ,Gastroenterology ,LGR5 ,Diseases of the digestive system. Gastroenterology ,Intestinal epithelium ,COX, cyclooxygenase ,Organoids ,ISC, intestinal stem cell ,Cancer research ,Tumor necrosis factor alpha ,Stem cell ,Crohn’s Disease - Abstract
Background & Aims Tumor necrosis factor alpha (TNFα) is considered a major tissue damage-promoting effector in Crohn’s disease (CD) pathogenesis. Patient-derived intestinal organoid (enteroid) recapitulates the disease-specific characteristics of the intestinal epithelium. This study aimed to evaluate the intestinal epithelial responses to TNFα in enteroids derived from healthy controls and compare them with those of CD patient-derived enteroids. Methods Human enteroids derived from patients with CD and controls were treated with TNFα (30 ng/mL), and cell viability and gene expression patterns were evaluated. Results TNFα induced MLKL-mediated necroptotic cell death, which was more pronounced in CD patient-derived enteroids than in control enteroids. Immunohistochemistry and RNA sequencing revealed that treatment with TNFα caused expansion of the intestinal stem cell (ISC) populations. However, expanded ISC subpopulations differed in control and CD patient-derived enteroids, with LGR5+ active ISCs in control enteroids and reserve ISCs, such as BMI1+ cells, in CD patient-derived enteroids. In single-cell RNA sequencing, LGR5+ ISC-enriched cell cluster showed strong expression of TNFRSF1B (TNFR2) and cyclooxygenase-prostaglandin E2 (PGE2) activation. In TNFα-treated CD patient-derived enteroids, exogenous PGE2 (10 nmol/L) induced the expansion of the LGR5+ ISC population and improved organoid-forming efficiency, viability, and wound healing. Conclusions TNFα increases necroptosis of differentiated cells and induces the expansion of LGR5+ ISCs. In CD patient-derived enteroids, TNFα causes LGR5+ stem cell dysfunction (expansion failure), and exogenous PGE2 treatment restored the functions of LGR5+ stem cells. Therefore, PGE2 can be used to promote mucosal healing in patients with CD., Graphical abstract
- Published
- 2022
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