Molecular Profile of Barrett's Esophagus and Gastroesophageal Reflux Disease in the Development of Translational Physiological and Pharmacological Studies.

Edyta Korbut,Marcin Surmiak,Dagmara Wójcik,Mateusz Wierdak,Jerzy Hankus,Katarzyna Magierowska,Tomasz Brzozowski,Marcin Magierowski,Maikel P Peppelenbosch,Vincent T Janmaat

doi:10.3390/ijms21176436

Edyta Korbut, Marcin Surmiak + Show 8 more

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https://doi.org/10.3390/ijms21176436

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Abstract

Barrett’s esophagus (BE) is a premalignant condition caused by gastroesophageal reflux disease (GERD), where physiological squamous epithelium is replaced by columnar epithelium. Several in vivo and in vitro BE models were developed with questionable translational relevance when implemented separately. Therefore, we aimed to screen Gene Expression Omnibus 2R (GEO2R) databases to establish whether clinical BE molecular profile was comparable with animal and optimized human esophageal squamous cell lines-based in vitro models. The GEO2R tool and selected databases were used to establish human BE molecular profile. BE-specific mRNAs in human esophageal cell lines (Het-1A and EPC2) were determined after one, three and/or six-day treatment with acidified medium (pH 5.0) and/or 50 and 100 µM bile mixture (BM). Wistar rats underwent microsurgical procedures to generate esophagogastroduodenal anastomosis (EGDA) leading to BE. BE-specific genes (keratin (KRT)1, KRT4, KRT5, KRT6A, KRT13, KRT14, KRT15, KRT16, KRT23, KRT24, KRT7, KRT8, KRT18, KRT20, trefoil factor (TFF)1, TFF2, TFF3, villin (VIL)1, mucin (MUC)2, MUC3A/B, MUC5B, MUC6 and MUC13) mRNA expression was assessed by real-time PCR. Pro/anti-inflammatory factors (interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, tumor necrosis factor α, interferon γ, granulocyte-macrophage colony-stimulating factor) serum concentration was assessed by a Luminex assay. Expression profile in vivo reflected about 45% of clinical BE with accompanied inflammatory response. Six-day treatment with 100 µM BM (pH 5.0) altered gene expression in vitro reflecting in 73% human BE profile and making this the most reliable in vitro tool taking into account two tested cell lines. Our optimized and established combined in vitro and in vivo BE models can improve further physiological and pharmacological studies testing pathomechanisms and novel therapeutic targets of this disorder.

Highlights

Barrett’s esophagus (BE) is a complex, genetically predisposed, premalignant condition of the distal esophagus characterized as replacement of the esophageal squamous epithelium into an intestinal-type columnar epithelium with a crypt-like architecture [1,2,3]
Expression of mRNA for squamous epithelium-specific genes such as KRT1, KRT4, KRT5, KRT6A-C, KRT13, KRT14, KRT15, KRT16, KRT23, KRT24 was significantly decreased in human Barrett’s esophagus biopsies as compared with expression of these specific genes in samples collected from normal squamous epithelium and observed in at least one out of three analyzed databases (p < 0.05, Table 1)
Expression of mRNA for columnar and intestinal epithelium-specific genes such as KRT7, KRT8, KRT18, KRT20, TFF1, TFF2, TFF3, VIL1, MUC2, MUC3A/B, MUC5B, MUC6, MUC13 was significantly upregulated in human BE biopsies when compared with gene expression in samples collected from normal squamous epithelium in at least one out of three analyzed databases (p < 0.05, Table 1)

Summary

Introduction

Barrett’s esophagus (BE) is a complex, genetically predisposed, premalignant condition of the distal esophagus characterized as replacement of the esophageal squamous epithelium into an intestinal-type columnar epithelium with a crypt-like architecture [1,2,3]. It has been confirmed that chronic gastroesophageal reflux disease (GERD) is one of the most important etiological component of BE [4,5]. BE and GERD are closely associated with a high risk of developing esophageal adenocarcinoma (EAC). The pattern of reflux is a significant factor that may influence the progression of BE towards advanced precancerous changes [6,7]. In contrast to the physiological esophageal epithelium, the BE development results in alternation of several individual molecular markers and signaling pathways. These changes include variety of mucins (MUC), mucin-associated trefoil factor family (TFF) peptides, and villin (VIL) [8,9,10]. The physiological and pathophysiological aspects still require further investigation, especially in the context of the implementation of novel non-invasive methods of treatment of this disorder

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