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Proton pump inhibitors modulate the barrier dysfunction caused by IL-13 in the epithelium in eosinophilic esophagitis

ABSTRACT: Eosinophilic esophagitis (EoE) is a chronic immune-mediated disorder triggered by specific food antigens, characterized by eosinophil-rich multicellular inflammation and structural changes in the epithelium. Treatment options for EoE include dietary therapy, pharmacological therapy, or a combination of both, chosen based on the patient's preference and clinical progression. Pharmacological options include proton pump inhibitors (PPIs), corticosteroids, and biologic therapy. Although PPI therapy is used for EoE management, its underlying mechanism of action remains unclear. To investigate the effects of omeprazole, an orally bioavailable PPI commonly used for EoE therapy, on the dynamics of esophageal epithelial barrier function and inflammation, we employed air-liquid interface (ALI) culture. We examined how omeprazole affects gene expression changes caused by IL-13 treatment. ALI cultures were treated with 100 ng/ml of IL-13 and/or 50 µM of acid-activated omeprazole for 96 hours, and bulk RNA sequencing was performed to analyze the epithelial-specific transcriptomes of IL-13 and/or omeprazole-treated ALI. Our findings suggest potential gene interactions where omeprazole may mitigate transcriptional changes induced by IL-13, indicating that omeprazole may attenuate IL-13 mediated epithelial dysfunction relevant to EoE pathophysiology by modulating pathways associated with inflammation, tissue repair, and cell-cell communication.

ORGANISM(S): Homo sapiens

PROVIDER: GSE273039 | GEO | 2024/09/13

REPOSITORIES: GEO

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Project description:Introduction Disruption of the epithelial barrier can induce eosinophilic esophagitis (EoE), a TH2-mediated food- and aeroallergen associated chronic inflammatory disease 1, 2. The expression of IL-20 subfamily cytokines, IL-19, IL-20 and IL-24, is increased in TH2-mediated diseases, yet their function in EoE is unknown. Methods Combining RNA-sequencing (RNA-seq) and proteome analysis, we have examined the effects of the IL-20 subfamily on esophageal epithelial cells using patient-derived organoids and an experimental EoE mouse model. Results When stimulated with IL-20 subfamily cytokines patient-derived esophageal organoids showed decreased expression of Filaggrin (FLG) and Filaggrin 2 (FLG2) responsible for epithelial cornification. In agreement, EoE patients with active inflammation showed elevated IL-20 subfamily cytokines and decreased FLG and FLG2 expression. Topical corticosteroid treatment in EoE patients restored filaggrin expression, while reducing IL-20 subfamily cytokines. Abolishment of IL-20 subfamily signalling in Il20R2-/- animals attenuated experimental EoE in an OVA-induced mouse model. In the esophageal keratinocyte cell line, KYSE-180, we identified IL-20 subfamily-induced STAT3 and MAPK (ERK1/2) pathway activation. However, Stat3fl/flKrt5cre mice with an epithelium specific deletion of Stat3 developed exacerbated experimental EoE with a pronounced decrease of Flg2. In line, combined stimulation of patient-derived esophageal organoids with IL-20 subfamily cytokines and pharmacological inhibition of STAT3 resulted in aggravated decrease of FLG and FLG2. In addition, pharmacological STAT3 inhibition resulted in reduced transepithelial electrical resistance (TEER) and increased macromolecular flux in patient-derived air liquid interface (ALI) cultures. Whereas, inhibition of the MAPK (ERK1/2) pathway hampered IL-20 subfamily induced TEER reduction and paracellular flux increment. Finally, MAPK (ERK1/2) inhibitor treatment reduced infiltrating CD45+ immune cells in the esophagus and alleviated experimental EoE in mice. Conclusion We discovered a novel regulatory function of the IL-20 subfamily for epithelial barrier integrity. Aberrant IL-20 subfamily signaling disturbs the epithelial barrier function, while abrogation of IL-20 subfamily and ERK1/2 signaling restores epithelial integrity and attenuates experimental EoE. Therefore, we propose that targeting the IL-20 subfamily pathway could present a novel therapeutic strategy for EoE treatment.

Project description:Background There is emerging evidence that the risk of inflammatory diseases in childhood such as eosinophilic esophagitis (EoE) is influenced by in-utero events, as prenatal complications increase the risk for the diseases. Objective We propose that the exposure of esophageal epithelium to amniotic fluid (AF) leads to alterations in its differentiation program and responsiveness to pro-allergic stimuli. Methods AF was obtained from the biorepository of the Genomic and Proteomic Network for Preterm Birth Research study. The cytokine/chemokine composition of AF was evaluated by a broad multiplex assay. Western blot was performed to interrogate signaling events following exposure of the esophageal epithelial cells to AF. A 3D spheroid model of esophageal epithelial differentiation was established and analyzed through bulk RNA sequencing and immunofluorescence. Effects of AF on esophageal epithelial differentiation and IL-13 response were assessed by RT-PCR. Results Multiplex analysis of AF revealed an expression of 17 cytokines with an average expression of ~ 70 – 3000 pg/ml. The highest expression was observed for MIP1 and IL1R, whereas IL-3, IL-4, IL-7, and TSLP were not detected. Several pro-inflammatory cytokines including TNF and IL-12 were highly expressed in the AF from women who underwent preterm birth. In contrast, the expression of EGF was the highest in the term birth samples. Exposure of esophageal epithelial cells to AF resulted in transient phosphorylation of ERK1/2 kinase and the transcription of early response genes including EGR1 and EGR2, highlighting the direct impact of AF on esophageal epithelial cells. AF modified the esophageal epithelial differentiation program, as evidenced by alterations in the expression of differentiation markers (ex. CRNN, KRT78, and SLURP1) in the 3D spheroids model. Moreover, AF enhanced the transcriptional response of genes associated with the IL-13 response, including CCL26 and CAPN14. Notably, the upregulation of CAPN14, encoded by the major EoE genetic susceptibility locus, was observed following exposure of epithelial cells to preterm AF samples independently of IL-13. Conclusions Collectively, our findings uncover an interplay between AF components and esophageal epithelial cells, emphasizing the potential relevance of AF in the pathogenesis of esophageal disorders such as EoE.

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Proton pump inhibitors modulate the barrier dysfunction caused by IL-13 in the epithelium in eosinophilic esophagitis

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