Interleukin-28A induces epithelial barrier dysfunction in CD patient-derived intestinal organoids

P. Xu*, H. Becker, M. Elizalde, M. Pierik, A. Masclee, D. Jonkers

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Intestinal barrier dysfunction is a pathogenic hallmark in Crohn?s disease (CD). Identifying key players that regulate intestinal barrier may provide novel leads for therapeutic intervention. Interleukin-28A (IL-28A) is a newly identified IL-10/interferon cytokine family member, with its most implicated function being antiviral and anti-proliferative properties. However, the role and underlying mechanisms of IL-28A in the regulation of epithelial barrier in CD remain so far unexplored. IL-28A levels were measured in the plasma and biopsies of CD patients and healthy subjects. CD patient-derived intestinal organoids were characterized by differentiation gene markers and then exposed to TNF-a, IFN-c, IL-113 or LPS, or IL-28A with or without GLPG0634 (filgotinib). Epithelial permeability was assessed by FITC-D4 flux. Expression of junctional components was analyzed by qRT-PCR, immunofluorescence staining, or Western blotting. JAK-STAT activity was analyzed by Western blotting. IL-28A levels were significantly increased in the plasma and biopsies from active patients with CD as compared with healthy subjects. IL-28A and its receptor complex IL-28AR/IL-10R2 were detected in CD patient-derived intestinal organoids and showed a selective response to IFN-c exposure. IL-28A triggered epithelial barrier disruption and accompanied by reduced ZO-1 and E-cadherin expression. This effect was mediated by JAK-STAT1 pathway. Pre-incubation with the JAK1 inhibitor filgotinib ameliorated the barrier dysfunction induced by IL-28A. These results identified IL-28A as a novel regulator of epithelial barrier function and could be a putative target for CD treatment. We provide novel basic evidence that restoring intestinal barrier is a potential mechanism that contributes to the clinical benefits of JAK1 inhibitor in patients with CD.NEW & NOTEWORTHY IL-28A levels were significantly increased in the plasma and biopsies from active patients with CD as compared with healthy subjects. IFN-c exposure stimulated IL-28A expression in intestinal organoids. Partially mimicking the effect of IFN-c, IL-28A impaired epithelial barrier function and disrupted junctional components through the activation of JAKSTAT1 signaling, whereas JAK1 inhibitor ameliorated the above-mentioned effects of IL-28A. These findings highlight the newly identified cytokine IL-28A as a novel contributor to CD pathogenesis and could be a putative target for CD treatment. We also provide new evidence for potential applications of JAK inhibition in CD therapy.
Original languageEnglish
Pages (from-to)G689-G699
Number of pages11
JournalAmerican Journal of Physiology-Gastrointestinal and Liver Physiology
Volume320
Issue number5
DOIs
Publication statusPublished - 1 May 2021

Keywords

  • ALPHA
  • CELLS
  • Crohn?s disease
  • EXPANSION
  • EXPRESSION
  • IL-29
  • INCREASES
  • JAK1 inhibitor
  • LOCI
  • NUMBER
  • PERMEABILITY
  • SIGNALS
  • epithelial barrier dysfunction
  • interleukin-28A
  • intestinal organoids
  • APOPTOSIS

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