IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model

Bailey Zwarycz, Adam D. Gracz, Kristina R. Rivera, Ian A. Williamson, Leigh A. Samsa, Joshua D Starmer, Michael A. Daniele, Luisa Salter-Cid, Qihong Zhao, Scott T Magness

Research output: Contribution to journalArticle

Abstract

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.

LanguageEnglish (US)
Pages1-17
Number of pages17
JournalCMGH
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2019

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Organoids
Inflammatory Bowel Diseases
Stem Cells
Epithelial Cells
Crohn Disease
interleukin-22
Interleukin-6
Cytokines
RNA Sequence Analysis
Ileum

Keywords

  • Inflammatory Bowel Disease
  • Interleukin-22
  • Intestinal Stem Cells

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

Cite this

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, Joshua D; Daniele, Michael A.; Salter-Cid, Luisa; Zhao, Qihong; Magness, Scott T.

In: CMGH, Vol. 7, No. 1, 01.01.2019, p. 1-17.

Research output: Contribution to journalArticle

Zwarycz, B, Gracz, AD, Rivera, KR, Williamson, IA, Samsa, LA, Starmer, JD, Daniele, MA, Salter-Cid, L, Zhao, Q & Magness, ST 2019, 'IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model' CMGH, vol. 7, no. 1, pp. 1-17. https://doi.org/10.1016/j.jcmgh.2018.06.008
Zwarycz B, Gracz AD, Rivera KR, Williamson IA, Samsa LA, Starmer JD et al. IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model. CMGH. 2019 Jan 1;7(1):1-17. https://doi.org/10.1016/j.jcmgh.2018.06.008
Zwarycz, Bailey ; Gracz, Adam D. ; Rivera, Kristina R. ; Williamson, Ian A. ; Samsa, Leigh A. ; Starmer, Joshua D ; Daniele, Michael A. ; Salter-Cid, Luisa ; Zhao, Qihong ; Magness, Scott T. / IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model. In: CMGH. 2019 ; Vol. 7, No. 1. pp. 1-17.
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T1 - IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model

AU - Zwarycz, Bailey

AU - Gracz, Adam D.

AU - Rivera, Kristina R.

AU - Williamson, Ian A.

AU - Samsa, Leigh A.

AU - Starmer, Joshua D

AU - Daniele, Michael A.

AU - Salter-Cid, Luisa

AU - Zhao, Qihong

AU - Magness, Scott T

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Y1 - 2019/1/1

N2 - 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.

AB - 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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