TNFα Induces LGR5+ Stem Cell Dysfunction In Patients With Crohn’s Disease

Autor: Chnasu Lee, Woong-Yang Park, Sung Noh Hong, Minae An, Je-Gun Joung, Dong Kyung Chang, Young-Ho Kim
Rok vydání: 2022
Předmět:
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
Zdroj: Cellular and Molecular Gastroenterology and Hepatology
Cellular and Molecular Gastroenterology and Hepatology, Vol 13, Iss 3, Pp 789-808 (2022)
ISSN: 2352-345X
DOI: 10.1016/j.jcmgh.2021.10.010
Popis: 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
Databáze: OpenAIRE
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