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Gut Dec 1960A study of 87 cases of carcinoma involving the gastro-oesophageal region included seven cases of adenocarcinoma confined to the anatomical limits of the oesophagus. Six...
A study of 87 cases of carcinoma involving the gastro-oesophageal region included seven cases of adenocarcinoma confined to the anatomical limits of the oesophagus. Six of these intraoesophageal adenocarcinomas appear to have arisen in "oesophagi lined by columnar epithelium". The seventh, a carcinoma of double histological pattern associated with a sliding hiatus hernia, showed a complex mixture of epithelial structures in the lower oesophagus. The histogenesis of intraoesophageal adenocarcinoma is discussed, and it is stressed that its prognosis may be relatively favourable.
Topics: Adenocarcinoma; Carcinoma; Esophageal Neoplasms; Female; Hernia, Hiatal; Humans; Medical Records
PubMed: 13723344
DOI: 10.1136/gut.1.4.351 -
Gut Mar 2002Minichromosome maintenance (Mcm) proteins are essential for eukaryotic DNA replication, and their expression implies potential for cell proliferation. Expression is...
BACKGROUND
Minichromosome maintenance (Mcm) proteins are essential for eukaryotic DNA replication, and their expression implies potential for cell proliferation. Expression is dysregulated in dysplastic states but data for oesophageal squamous mucosa and Barrett's mucosa have not been published.
AIM
To test the hypothesis that Mcm proteins are downregulated together with the proliferation marker Ki-67 in differentiating epithelial compartments of non-dysplastic squamous and Barrett's epithelium, and that this process does not occur in dysplastic mucosae. METHODS AND CASES: Forty five patients with Barrett's oesophagus included 20 with glandular dysplasia (10 low grade, eight high grade, two both, and four with invasive adenocarcinoma). Twenty five other patients included 12 with oesophageal squamous dysplasia (three low grade, six high grade, three both, and four with invasive squamous carcinoma). Formalin fixed paraffin embedded tissue sections from biopsy series and resections were immunostained using antibodies to Mcm2, Mcm5, and Ki-67. Percentage of nuclei positive for Mcm2, Mcm5, and Ki-67 was estimated and scored from 0 to 6 as: 0, none +; 1, <10%+; 2, 10-30%+; 3, 30-70%+; 4, 70-90%+; 5, >90%+; 6, all+. Four separate epithelial strata were scored: in squamous epithelium the basal layer and thirds to the surface, in Barrett's mucosa the luminal surface, upper and lower crypt, and deep glands.
RESULTS
In non-dysplastic squamous epithelium and Barrett's mucosa, high level expression of Mcm2, Mcm5, and Ki-67 proteins was largely confined to the proliferative compartments and downregulated in differentiated compartments. Expression persisted up to the mucosal surface in dysplastic squamous epithelium and Barrett's mucosa.
CONCLUSIONS
Persistent expression of Mcm2, Mcm5, and Ki-67 proteins in luminal compartments of dysplastic oesophageal squamous epithelium and dysplastic Barrett's mucosa may be diagnostic markers and imply disruption of cell cycle control and differentiation in these dysplastic epithelia.
Topics: Barrett Esophagus; Biomarkers, Tumor; Cell Cycle Proteins; Cell Division; DNA-Binding Proteins; Down-Regulation; Esophageal Neoplasms; Humans; Ki-67 Antigen; Minichromosome Maintenance Complex Component 2; Neoplasm Proteins; Nuclear Proteins; Precancerous Conditions; Reproducibility of Results; Schizosaccharomyces pombe Proteins
PubMed: 11839717
DOI: 10.1136/gut.50.3.373 -
Gut Jul 2017The influence of eosinophilic oesophagitis (EoE)-associated inflammation upon oesophageal epithelial biology remains poorly understood. We investigated the functional...
OBJECTIVE
The influence of eosinophilic oesophagitis (EoE)-associated inflammation upon oesophageal epithelial biology remains poorly understood. We investigated the functional role of autophagy in oesophageal epithelial cells (keratinocytes) exposed to the inflammatory EoE milieu.
DESIGN
Functional consequences of genetic or pharmacological autophagy inhibition were assessed in endoscopic oesophageal biopsies, human oesophageal keratinocytes, single cell-derived ex vivo murine oesophageal organoids as well as a murine model recapitulating EoE-like inflammation and basal cell hyperplasia. Gene expression, morphological and functional characterisation of autophagy and oxidative stress were performed by transmission electron microscopy, immunostaining, immunoblotting, live cell imaging and flow cytometry.
RESULTS
EoE-relevant inflammatory conditions promoted autophagy and basal cell hyperplasia in three independent murine EoE models and oesophageal organoids. Inhibition of autophagic flux via chloroquine treatment augmented basal cell hyperplasia in these model systems. Oesophageal keratinocytes stimulated with EoE-relevant cytokines, including tumour necrosis factor-α and interleukin-13 exhibited activation of autophagic flux in a reactive oxygen species-dependent manner. Autophagy inhibition via chloroquine treatment or depletion of Beclin-1 or ATG-7, augmented oxidative stress induced by EoE-relevant stimuli in murine EoE, oesophageal organoids and human oesophageal keratinocytes. Oesophageal epithelia of paediatric EoE patients with active inflammation displayed increased autophagic vesicle content compared with normal and EoE remission subjects. Functional flow cytometric analysis revealed autophagic flux in human oesophageal biopsies.
CONCLUSIONS
Our findings reveal for the first time that autophagy may function as a cytoprotective mechanism to maintain epithelial redox balance and homeostasis under EoE inflammation-associated stress, providing mechanistic insights into the role of autophagy in EoE pathogenesis.
Topics: Animals; Autophagy; Chloroquine; Cytokines; Eosinophilic Esophagitis; Eosinophils; Epithelium; Esophagoscopy; Esophagus; Humans; Keratinocytes; Mice; Models, Animal; Oxidative Stress
PubMed: 26884425
DOI: 10.1136/gutjnl-2015-310341 -
Gut Apr 1975
Review
Topics: Aldosterone; Biological Transport, Active; Carbenoxolone; Colitis; Colon; Dinitrophenols; Electrodes; Electrophysiology; Epithelium; Humans; Intestinal Mucosa; Ouabain; Proctitis; Sodium; Spironolactone
PubMed: 1093951
DOI: 10.1136/gut.16.4.315 -
Gut Mar 2005Barrett's oesophagus is a premalignant condition that predisposes to the development of oesophageal adenocarcinoma. It is detected on endoscopy and confirmed... (Review)
Review
Barrett's oesophagus is a premalignant condition that predisposes to the development of oesophageal adenocarcinoma. It is detected on endoscopy and confirmed histologically by the presence in the lower oesophagus of a metaplastic mucosa, the so-called specialised epithelium, which resembles incomplete intestinal metaplasia in the stomach. These similarities with incomplete intestinal metaplasia are present on histology, mucin histochemistry, and immunohistochemistry with various differentiation markers (cytokeratins and MUC antigens). On morphology, the carcinogenetic process of Barrett's mucosa progresses through increasing grades of epithelial dysplasia. Dysplasia, a synonym of intraepithelial neoplasia, is the only marker that can be used at the present time to delineate a population of patients at high risk of cancer. Among the numerous molecular events that have been shown to play a role in the neoplastic transformation of Barrett's mucosa, only changes in DNA ploidy, increased proliferation, and alterations of the p53 gene have been suggested to be of potential help in the surveillance of patients.
Topics: Aneuploidy; Barrett Esophagus; Cell Transformation, Neoplastic; DNA, Neoplasm; Esophageal Neoplasms; Esophagus; Genes, p53; Humans; Immunohistochemistry; Mucins; Mucous Membrane; Neoplasms, Glandular and Epithelial; Precancerous Conditions; Risk Factors
PubMed: 15711008
DOI: 10.1136/gut.2004.041525 -
Gut Aug 2018Nuclear receptors are known to regulate both immune and barrier functions in the GI tract. The nuclear orphan receptor NR2F6 has been shown to suppress the expression of...
OBJECTIVE
Nuclear receptors are known to regulate both immune and barrier functions in the GI tract. The nuclear orphan receptor NR2F6 has been shown to suppress the expression of proinflammatory cytokines in T lymphocytes. gene expression is reduced in patients with IBS or UC, but its functional role and tissue dependency in healthy and inflamed gut have not yet been investigated.
DESIGN
Intestinal inflammation was induced in wild-type, -deficient, -deficient or bone marrow-reconstituted mice by administration of chemical (dextran sodium sulfate (DSS)) and immunogenic (T cell transfer) triggers. Disease phenotypes were investigated by survival, body weight, colon length and analysis of immune cell infiltrates. Additionally, histology, intestinal permeability, tight junction proteins, bacterial fluorescence in situ hybridisation, apoptosis, cell proliferation and mucus production were investigated.
RESULTS
-deficient mice were highly susceptible to DSS-induced colitis characterised by enhanced weight loss, increased colonic tissue destruction and immune cell infiltration together with enhanced intestinal permeability and reduced expression. T cell transfer colitis and bone marrow reconstitution experiments demonstrated that disease susceptibility was not dependent on the expression of in the immune compartment but on the protective role of NR2F6 in the intestinal epithelium. Mechanistically, we show that NR2F6 binds to a consensus sequence at -2 kb of the promoter and transactivates expression. Loss of NR2F6 alters intestinal permeability and results in spontaneous late-onset colitis in -deficient mice.
CONCLUSION
We have for the first time identified a fundamental and non-redundant role of NR2F6 in protecting gut barrier homeostasis.
Topics: Animals; COUP Transcription Factors; Colitis; Dextran Sulfate; Disease Models, Animal; Intestinal Mucosa; Mice; Mucin-2; Repressor Proteins; Tight Junction Proteins
PubMed: 28779026
DOI: 10.1136/gutjnl-2016-313466 -
Gut May 2014As a major cellular defence mechanism, the Nrf2/Keap1 pathway regulates expression of genes involved in detoxification and stress response. Here we hypothesise that Nrf2...
OBJECTIVE
As a major cellular defence mechanism, the Nrf2/Keap1 pathway regulates expression of genes involved in detoxification and stress response. Here we hypothesise that Nrf2 is involved in oesophageal barrier function and plays a protective role against gastro-oesophageal reflux disease (GERD).
DESIGN
Human oesophageal biopsy samples, mouse surgical models and Nrf2(-/-) mice were used to assess the role of the Nrf2/Keap1 pathway in oesophageal barrier function. Trans-epithelial electrical resistance (TEER) was measured with mini-Ussing chambers. HE staining and transmission electron microscopy were used to examine tissue morphology, while gene microarray, immunohistochemistry, western blotting and chromatin immunoprecipitation (ChIP) analysis were used to assess gene expression.
RESULTS
Nrf2 was expressed in normal oesophageal epithelium and activated in GERD of both humans and mice. Nrf2 deficiency and gastro-oesophageal reflux in mice, alone or in combination, reduced TEER and increased intercellular space in oesophageal epithelium. Nrf2 target genes and gene sets associated with oxidoreductase activity, mitochondrial biogenesis and energy production were downregulated in the oesophageal epithelium of Nrf2(-/-) mice. Consistent with the antioxidative function of Nrf2, a DNA oxidative damage marker (8OHdG) dramatically increased in oesophageal epithelial cells of Nrf2(-/-) mice compared with those of wild-type mice. Interestingly, ATP biogenesis, Cox IV (a mitochondrial protein) and Claudin 4 (Cldn4) expression were downregulated in the oesophageal epithelium of Nrf2(-/-) mice, suggesting that energy-dependent tight junction integrity was subject to Nrf2 regulation. ChIP analysis confirmed the binding of Nrf2 to Cldn4 promoter.
CONCLUSIONS
Nrf2 deficiency impairs oesophageal barrier function through disrupting energy-dependent tight junction.
Topics: Adaptor Proteins, Signal Transducing; Animals; Biomarkers; Blotting, Western; Chromatin Immunoprecipitation; Cytoskeletal Proteins; Down-Regulation; Electric Impedance; Esophagus; Gastroesophageal Reflux; Humans; Immunohistochemistry; Kelch-Like ECH-Associated Protein 1; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Microscopy, Electron, Transmission; Mucous Membrane; NF-E2-Related Factor 2; Oligonucleotide Array Sequence Analysis; Oxidative Stress; Permeability; Tight Junctions
PubMed: 23676441
DOI: 10.1136/gutjnl-2012-303731 -
Gut May 2001Colonic epithelium is involved in the regulation of intestinal function and mucosal immune responses, and its function is altered in inflammatory bowel disease (IBD)....
BACKGROUND AND AIMS
Colonic epithelium is involved in the regulation of intestinal function and mucosal immune responses, and its function is altered in inflammatory bowel disease (IBD). However, a comprehensive analysis of the genetic alterations in inflamed colonic epithelium is not available at present. The aim of our study was to detect genes that are preferentially expressed in inflamed colonic epithelia and clarify the biochemical responses of epithelial cells in inflamed colonic mucosa.
METHODS
cDNA representation difference analysis was used to identify candidate genes selectively expressed in inflamed colonic epithelia. Selective expression of these genes in the epithelium of inflamed colonic mucosa, including IBD and non-IBD tissues, was examined by real time polymerase chain reaction and in situ hybridisation. The effect of cell confluence and inflammatory mediators on Reg 1alpha gene expression was examined using a colon cancer cell line (HT29).
RESULTS
We identified seven candidate genes that were presumed to be upregulated in the inflamed colonic epithelium. Of these, Reg 1alpha and GW112 were the dominant species and expression of these genes was confined to the crypt epithelium. In vitro studies using a colonic epithelial cell line suggested that cell confluence regulates Reg 1alpha gene expression.
CONCLUSIONS
Selective expression of Reg 1alpha and GW112 genes in the crypt epithelium of inflamed colonic mucosa suggests the important regulatory functions of these genes.
Topics: Adult; Aged; Annexin A1; Case-Control Studies; Colonic Neoplasms; Cytokines; DNA, Complementary; Female; Gene Expression; Genes, Regulator; Humans; In Situ Hybridization; Inflammatory Bowel Diseases; Intestinal Mucosa; Male; Middle Aged; Polymerase Chain Reaction; Sequence Analysis, DNA; Tumor Cells, Cultured; Up-Regulation
PubMed: 11302958
DOI: 10.1136/gut.48.5.623 -
Gut Apr 2004Chronic stress affects the course of inflammatory bowel disease and experimental colitis, and may also initiate intestinal inflammation in rats.
BACKGROUND
Chronic stress affects the course of inflammatory bowel disease and experimental colitis, and may also initiate intestinal inflammation in rats.
AIM
To investigate the effects of stress on the M cell containing follicle associated epithelium, specialised in antigen uptake.
SUBJECTS AND METHODS
Wistar rats were submitted to acute water avoidance stress for one hour or chronic water avoidance stress for 1 hour/day for 10 consecutive days. Permeability to (51)Cr-EDTA, horseradish peroxidase, and chemically killed Escherichia coli K-12 was studied in both villus and follicle associated epithelium in Ussing chambers. Segments were further examined by light, electron, and confocal microscopy.
RESULTS
Acute stress increased horseradish peroxidase flux in villus as well as in follicle associated epithelium. Chronic stress further increased permeability to horseradish peroxidase in villus and follicle associated epithelium, in the latter by almost fourfold. Moreover, chronic stress induced over 30 times increased E coli passage in follicle associated epithelium whereas there was no significant increase in villus epithelium. Bacterial uptake was confirmed by confocal microscopy showing fluorescent bacteria penetrating and passing through the epithelial surface.
CONCLUSIONS
These results show that the barrier function of follicle associated epithelium can be modulated, and that chronic stress enhances the uptake of luminal antigens and bacteria via the follicle associated epithelium. This can increase antigen exposure in Peyer's patches thereby having implications in the initiation of proinflammatory immune responses within the intestinal mucosa.
Topics: Acute Disease; Animals; Antigens; Bacterial Translocation; Chronic Disease; Electric Conductivity; Escherichia coli; Intestinal Absorption; Intestinal Mucosa; Male; Permeability; Peyer's Patches; Rats; Rats, Wistar; Stress, Psychological
PubMed: 15016742
DOI: 10.1136/gut.2003.028506 -
Gut Jan 2019Human intestinal epithelial organoids (IEOs) are increasingly being recognised as a highly promising translational research tool. However, our understanding of their...
OBJECTIVE
Human intestinal epithelial organoids (IEOs) are increasingly being recognised as a highly promising translational research tool. However, our understanding of their epigenetic molecular characteristics and behaviour in culture remains limited.
DESIGN
We performed genome-wide DNA methylation and transcriptomic profiling of human IEOs derived from paediatric/adult and fetal small and large bowel as well as matching purified human gut epithelium. Furthermore, organoids were subjected to in vitro differentiation and genome editing using CRISPR/Cas9 technology.
RESULTS
We discovered stable epigenetic signatures which define regional differences in gut epithelial function, including induction of segment-specific genes during cellular differentiation. Established DNA methylation profiles were independent of cellular environment since organoids retained their regional DNA methylation over prolonged culture periods. In contrast to paediatric and adult organoids, fetal gut-derived organoids showed distinct dynamic changes of DNA methylation and gene expression in culture, indicative of an in vitro maturation. By applying CRISPR/Cas9 genome editing to fetal organoids, we demonstrate that this process is partly regulated by TET1, an enzyme involved in the DNA demethylation process. Lastly, generating IEOs from a child diagnosed with gastric heterotopia revealed persistent and distinct disease-associated DNA methylation differences, highlighting the use of organoids as disease-specific research models.
CONCLUSIONS
Our study demonstrates striking similarities of epigenetic signatures in mucosa-derived IEOs with matching primary epithelium. Moreover, these results suggest that intestinal stem cell-intrinsic DNA methylation patterns establish and maintain regional gut specification and are involved in early epithelial development and disease.
Topics: Cell Differentiation; Cells, Cultured; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Methylation; Epigenesis, Genetic; Epithelial Cells; Humans; Intestinal Mucosa; Organoids; Transcriptome
PubMed: 29141958
DOI: 10.1136/gutjnl-2017-314817