IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model.
Animals
Biomarkers
/ metabolism
Cell Lineage
/ drug effects
Cell Proliferation
/ drug effects
Cell Self Renewal
/ drug effects
Cellular Microenvironment
/ drug effects
Computer Simulation
Epithelial Cells
/ cytology
Ileum
/ cytology
Inflammatory Bowel Diseases
/ pathology
Interleukins
/ pharmacology
Mice, Inbred C57BL
Models, Biological
Organoids
/ cytology
Receptors, Interleukin
/ metabolism
Serum
/ metabolism
Stem Cells
/ cytology
Interleukin-22
BSA, bovine serum albumin
EGFP, enhanced green fluorescent protein
FACS, fluorescence-activated cell sorter
IBD, inflammatory bowel disease
IL, interleukin
IL22RA1, IL22 receptor A1
IL22TG, IL22 transgenic
ILC, innate lymphoid cell
ILC3, IL22-secreting lymphocyte
ISC, intestinal stem cell
Inflammatory Bowel Disease
Interleukin-22
Intestinal Stem Cells
OFE, organoid forming efficiency
STAT3, signal transducer and activator of transcription 3
TA, transit-amplifying
TBS, Tris-buffered saline
cDNA, complementary DNA
mRNA, messenger RNA
Journal
Cellular and molecular gastroenterology and hepatology
ISSN: 2352-345X
Titre abrégé: Cell Mol Gastroenterol Hepatol
Pays: United States
ID NLM: 101648302
Informations de publication
Date de publication:
2019
2019
Historique:
received:
07
09
2017
accepted:
25
06
2018
entrez:
27
10
2018
pubmed:
27
10
2018
medline:
30
4
2019
Statut:
epublish
Résumé
Crohn's disease is an inflammatory bowel disease that affects the ileum and is associated with increased cytokines. Although interleukin (IL)6, IL17, IL21, and IL22 are increased in Crohn's disease and are associated with disrupted epithelial regeneration, little is known about their effects on the intestinal stem cells (ISCs) that mediate tissue repair. We hypothesized that ILs may target ISCs and reduce ISC-driven epithelial renewal. A screen of IL6, IL17, IL21, or IL22 was performed on ileal mouse organoids. Computational modeling was used to predict microenvironment cytokine concentrations. Organoid size, survival, proliferation, and differentiation were characterized by morphometrics, quantitative reverse-transcription polymerase chain reaction, and immunostaining on whole organoids or isolated ISCs. ISC function was assayed using serial passaging to single cells followed by organoid quantification. Single-cell RNA sequencing was used to assess High IL22 levels caused decreased ileal organoid survival, however, resistant organoids grew larger and showed increased proliferation over controls. Increased IL22 limits ISC expansion in favor of increased TA progenitor cell expansion.
Sections du résumé
Background & Aims
Crohn's disease is an inflammatory bowel disease that affects the ileum and is associated with increased cytokines. Although interleukin (IL)6, IL17, IL21, and IL22 are increased in Crohn's disease and are associated with disrupted epithelial regeneration, little is known about their effects on the intestinal stem cells (ISCs) that mediate tissue repair. We hypothesized that ILs may target ISCs and reduce ISC-driven epithelial renewal.
Methods
A screen of IL6, IL17, IL21, or IL22 was performed on ileal mouse organoids. Computational modeling was used to predict microenvironment cytokine concentrations. Organoid size, survival, proliferation, and differentiation were characterized by morphometrics, quantitative reverse-transcription polymerase chain reaction, and immunostaining on whole organoids or isolated ISCs. ISC function was assayed using serial passaging to single cells followed by organoid quantification. Single-cell RNA sequencing was used to assess
Results
High IL22 levels caused decreased ileal organoid survival, however, resistant organoids grew larger and showed increased proliferation over controls.
Conclusions
Increased IL22 limits ISC expansion in favor of increased TA progenitor cell expansion.
Identifiants
pubmed: 30364840
doi: 10.1016/j.jcmgh.2018.06.008
pii: S2352-345X(18)30094-8
pmc: PMC6199238
pii:
doi:
Substances chimiques
Biomarkers
0
Interleukins
0
Receptors, Interleukin
0
interleukin-22 receptor
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1-17Subventions
Organisme : NIDDK NIH HHS
ID : F31 DK107137
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK034987
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK091427
Pays : United States
Commentaires et corrections
Type : CommentIn
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