Corticosteroids impair epithelial regeneration in immune-mediated intestinal damage
Viktor Arnhold, Winston Y. Chang, Suze A. Jansen, Govindarajan Thangavelu, Marco Calafiore, Paola Vinci, Ya-Yuan Fu, Takahiro Ito, Shuichiro Takashima, Anastasiya Egorova, Jason Kuttiyara, Adam Perlstein, Marliek van Hoesel, Chen Liu, Bruce R. Blazar, Caroline A. Lindemans, Alan M. Hanash
Viktor Arnhold, Winston Y. Chang, Suze A. Jansen, Govindarajan Thangavelu, Marco Calafiore, Paola Vinci, Ya-Yuan Fu, Takahiro Ito, Shuichiro Takashima, Anastasiya Egorova, Jason Kuttiyara, Adam Perlstein, Marliek van Hoesel, Chen Liu, Bruce R. Blazar, Caroline A. Lindemans, Alan M. Hanash
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Research Article Immunology

Corticosteroids impair epithelial regeneration in immune-mediated intestinal damage

Abstract

Corticosteroid treatment (CST) failure is associated with poor outcomes for patients with gastrointestinal (GI) graft-versus-host disease (GVHD). CST is intended to target the immune system, but the glucocorticoid receptor (GR) is widely expressed, including within the intestines, where its effects are poorly understood. Here, we report that corticosteroids (CS) directly targeted intestinal epithelium, potentially worsening immune-mediated GI damage. CS administered to mice in vivo and intestinal organoid cultures ex vivo reduced epithelial proliferation. Following irradiation, immediate CST mitigated GI damage but delayed treatment attenuated regeneration and exacerbated damage. In a murine steroid-refractory (SR) GVHD model, CST impaired epithelial regeneration, worsened crypt loss, and reduced intestinal stem cell (ISC) frequencies. CST also exacerbated immune-mediated damage in organoid cultures with SR, GR-deficient T cells or IFN-γ. These findings correlated with CS-dependent changes in apoptosis-related gene expression and STAT3-related epithelial proliferation. Conversely, IL-22 administration enhanced STAT3 activity and overcame CS-mediated attenuation of regeneration, reducing crypt loss and promoting ISC expansion in steroid-treated mice with GVHD. Therefore, CST has the potential to exacerbate GI damage if it fails to control the damage-inducing immune response, but this risk may be countered by strategies augmenting epithelial regeneration, thus providing a rationale for clinical approaches combining such tissue-targeted therapies with immunosuppression.

Authors

Viktor Arnhold, Winston Y. Chang, Suze A. Jansen, Govindarajan Thangavelu, Marco Calafiore, Paola Vinci, Ya-Yuan Fu, Takahiro Ito, Shuichiro Takashima, Anastasiya Egorova, Jason Kuttiyara, Adam Perlstein, Marliek van Hoesel, Chen Liu, Bruce R. Blazar, Caroline A. Lindemans, Alan M. Hanash

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Figure 4

Epithelial effects of CS treatment after irradiation are timing dependent.

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Epithelial effects of CS treatment after irradiation are timing dependen...
(A–D) WT B6 mice were treated with MP (2 mg/kg) or vehicle i.p. daily starting 24 hours after TBI. (B and C) Representative images, ileal crypt frequency, and height, 5 days after TBI (n = 21–25 sections per group). Scale bars: 50 μm. (D) Representative Olfm4 IHC staining and Olfm4+ cell frequencies 5 days after TBI (n = 25–27 sections per group). Scale bars: 50 μm. (E) GSEA of the MSigDB apoptosis gene set in SI epithelial cells from WT mice treated with DEX or vehicle. One analysis and a nominal P value are shown. (F and G) SI crypt cells were plated 4 hours prior to 4 Gy irradiation. Cultures were treated with MP (10 μM) 16 hours after irradiation. (F) Organoids were evaluated for frequency and size 3 days after irradiation (n = 3–4 wells per group). (G) Bcl2l1 and Bik expression was determined by RT-qPCR 48 hours after irradiation (n = 3 wells per group). (H–L) WT B6 animals were treated with MP (2 mg/kg) or vehicle i.p. daily, starting 72 hours after TBI. (I and J) Representative images and ileal crypt frequency and height (n = 15–21 sections per group), 7 days after TBI. Scale bars: 50 μm. (K) Representative Ki67 IHC images and data showing Ki67+ cell frequencies, 7 days after TBI (n = 19–21 sections per group). Scale bars: 50 μm. (L) RT-qPCR showing Ccna2 and Ccnb1 expression in enriched SI crypts, 7 days after TBI (n = 8 animals per group). (M) SI crypt cells were plated 4 hours prior to 4 Gy irradiation; cultures were treated with MP (10 μM) 3 days after irradiation. Seven days after irradiation, organoids were evaluated for frequency and size (n = 7–10 wells per group). *P < 0.05, **P < 0.01, and ***P < 0.001, by 2-tailed t test or 1-way ANOVA. Data are representative of at least 2 independent experiments or were combined from 2 independent experiments (A–D and H–L).