Cargando…
IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model
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 abou...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199238/ https://www.ncbi.nlm.nih.gov/pubmed/30364840 http://dx.doi.org/10.1016/j.jcmgh.2018.06.008 |
_version_ | 1783365100494651392 |
---|---|
author | Zwarycz, Bailey Gracz, Adam D. Rivera, Kristina R. Williamson, Ian A. Samsa, Leigh A. Starmer, Josh Daniele, Michael A. Salter-Cid, Luisa Zhao, Qihong Magness, Scott T. |
author_facet | Zwarycz, Bailey Gracz, Adam D. Rivera, Kristina R. Williamson, Ian A. Samsa, Leigh A. Starmer, Josh Daniele, Michael A. Salter-Cid, Luisa Zhao, Qihong Magness, Scott T. |
author_sort | Zwarycz, Bailey |
collection | PubMed |
description | 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 Il22ra1 expression patterns in ISCs and transit-amplifying (TA) progenitors. An IL22-transgenic mouse was used to confirm the impact of increased IL22 on proliferative cells in vivo. RESULTS: High IL22 levels caused decreased ileal organoid survival, however, resistant organoids grew larger and showed increased proliferation over controls. Il22ra1 was expressed on only a subset of ISCs and TA progenitors. IL22-treated ISCs did not show appreciable differentiation defects, but ISC biomarker expression and self-renewal–associated pathway activity was reduced and accompanied by an inhibition of ISC expansion. In vivo, chronically increased IL22 levels, similar to predicted microenvironment levels, showed increases in proliferative cells in the TA zone with no increase in ISCs. CONCLUSIONS: Increased IL22 limits ISC expansion in favor of increased TA progenitor cell expansion. |
format | Online Article Text |
id | pubmed-6199238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-61992382018-10-25 IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model Zwarycz, Bailey Gracz, Adam D. Rivera, Kristina R. Williamson, Ian A. Samsa, Leigh A. Starmer, Josh Daniele, Michael A. Salter-Cid, Luisa Zhao, Qihong Magness, Scott T. Cell Mol Gastroenterol Hepatol Original Research 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 Il22ra1 expression patterns in ISCs and transit-amplifying (TA) progenitors. An IL22-transgenic mouse was used to confirm the impact of increased IL22 on proliferative cells in vivo. RESULTS: High IL22 levels caused decreased ileal organoid survival, however, resistant organoids grew larger and showed increased proliferation over controls. Il22ra1 was expressed on only a subset of ISCs and TA progenitors. IL22-treated ISCs did not show appreciable differentiation defects, but ISC biomarker expression and self-renewal–associated pathway activity was reduced and accompanied by an inhibition of ISC expansion. In vivo, chronically increased IL22 levels, similar to predicted microenvironment levels, showed increases in proliferative cells in the TA zone with no increase in ISCs. CONCLUSIONS: Increased IL22 limits ISC expansion in favor of increased TA progenitor cell expansion. Elsevier 2018-07-04 /pmc/articles/PMC6199238/ /pubmed/30364840 http://dx.doi.org/10.1016/j.jcmgh.2018.06.008 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Research Zwarycz, Bailey Gracz, Adam D. Rivera, Kristina R. Williamson, Ian A. Samsa, Leigh A. Starmer, Josh Daniele, Michael A. Salter-Cid, Luisa Zhao, Qihong Magness, Scott T. IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title | IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title_full | IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title_fullStr | IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title_full_unstemmed | IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title_short | IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model |
title_sort | il22 inhibits epithelial stem cell expansion in an ileal organoid model |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199238/ https://www.ncbi.nlm.nih.gov/pubmed/30364840 http://dx.doi.org/10.1016/j.jcmgh.2018.06.008 |
work_keys_str_mv | AT zwaryczbailey il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT graczadamd il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT riverakristinar il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT williamsoniana il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT samsaleigha il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT starmerjosh il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT danielemichaela il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT saltercidluisa il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT zhaoqihong il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel AT magnessscottt il22inhibitsepithelialstemcellexpansioninanilealorganoidmodel |