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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...

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Autores principales: 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.
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
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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.
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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
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