ISG15/GRAIL1/CD3 axis influences survival of patients with esophageal adenocarcinoma.

Immunosuppression is a common feature of esophageal adenocarcinoma (EAC) and has been linked to poor overall survival (OS). We hypothesized that upstream factors might negatively influence CD3 levels and T cell activity, thus promoting immunosuppression and worse survival. We used clinical data and patient samples of those who progressed from Barrett's to dysplasia to EAC, investigated gene (RNA-Seq) and protein (tissue microarray) expression, and performed cell biology studies to delineate a pathway impacting CD3 protein stability that might influence EAC outcome. We showed that the loss of both CD3-ε expression and CD3+ T cell number correlated with worse OS in EAC. The gene related to anergy in lymphocytes isoform 1 (GRAIL1), which is the prominent isoform in EACs, degraded (ε, γ, δ) CD3s and inactivated T cells. In contrast, isoform 2 (GRAIL2), which is reduced in EACs, stabilized CD3s. Further, GRAIL1-mediated CD3 degradation was facilitated by interferon-stimulated gene 15 (ISG15), a ubiquitin-like protein. Consequently, the overexpression of a ligase-dead GRAIL1, ISG15 knockdown, or the overexpression of a conjugation-defective ISG15-leucine-arginine-glycine-glycine mutant could increase CD3 levels. Together, we identified an ISG15/GRAIL1/mutant p53 amplification loop negatively influencing CD3 levels and T cell activity, thus promoting immunosuppression in EAC.

Hypoxia-inducible factor-1α mediates reflux-induced epithelial-mesenchymal plasticity in Barrett's oesophagus patients.

INTRODUCTION: Epithelial-mesenchymal plasticity (EMP), the process through which epithelial cells acquire mesenchymal features, is needed for wound repair but also might contribute to cancer initiation. Earlier, in vitro studies showed that Barrett's cells exposed to acidic bile salt solutions (ABS) develop EMP. Now, we have (1) induced reflux oesophagitis in Barrett's oesophagus (BO) patients by stopping proton pump inhibitors (PPIs), (2) assessed their biopsies for EMP and (3) explored molecular pathways underlying reflux-induced EMP in BO cells and spheroids. METHODS: 15 BO patients had endoscopy with biopsies of Barrett's metaplasia while on PPIs, and 1 and 2 weeks after stopping PPIs; RNA-seq data were assessed for enrichments in hypoxia-inducible factors (HIFs), angiogenesis and EMP pathways. In BO biopsies, cell lines and spheroids, EMP features (motility) and markers (vascular endothelial growth factor (VEGF), ZEB1, miR-200a&b) were evaluated by morphology, migration assays, immunostaining and qPCR; HIF-1α was knocked down with siRNA or shRNA. RESULTS: At 1 and/or 2 weeks off PPIs, BO biopsies exhibited EMP features and markers, with significant enrichment for HIF-1α, angiogenesis and EMP pathways. In BO cells, ABS induced HIF-1α activation, which decreased miR-200a&b while increasing VEGF, ZEB1 and motility; HIF-1α knockdown blocked these effects. After ABS treatment, BO spheroids exhibited migratory protrusions showing nuclear HIF-1α, increased VEGF and decreased miR-200a&b. CONCLUSIONS: In BO patients, reflux oesophagitis induces EMP changes associated with increased HIF-1α signalling in Barrett's metaplasia. In Barrett's cells, ABS trigger EMP via HIF-1α signalling. Thus, HIF-1α appears to play a key role in mediating reflux-induced EMP that might contribute to cancer in BO. TRIAL REGISTRATION NUMBER: NCT02579460.

miRNA profiling of esophageal adenocarcinoma using transcriptome analysis.

Esophageal adenocarcinoma (EAC) occurs following a series of histological changes through epithelial-mesenchymal transition (EMT). A variable expression of normal and aberrant genes in the tissue can contribute to the development of EAC through the activation or inhibition of critical molecular signaling pathways. Gene expression is regulated by various regulatory factors, including transcription factors and microRNAs (miRs). The exact profile of miRs associated with the pathogenesis of EAC is largely unknown, though some candidate miRNAs have been reported in the literature. To identify the unique miR profile associated with EAC, we compared normal esophageal tissue to EAC tissue using bulk RNA sequencing. RNA sequence data was verified using qPCR of 18 selected genes. Fourteen were confirmed as being upregulated, which include CDH11, PCOLCE, SULF1, GJA4, LUM, CDH6, GNA12, F2RL2, CTSZ, TYROBP, and KDELR3 as well as the downregulation of UGT1A1. We then conducted Ingenuity Pathway Analysis (IPA) to analyze for novel miR-gene relationships through Causal Network Analysis and Upstream Regulator Analysis. We identified 46 miRs that were aberrantly expressed in EAC compared to control tissues. In EAC tissues, seven miRs were associated with activated networks, while 39 miRs were associated with inhibited networks. The miR-gene relationships identified provide novel insights into potentially oncogenic molecular pathways and genes associated with carcinogenesis in esophageal tissue. Our results revealed a distinct miR profile associated with dysregulated genes. The miRs and genes identified in this study may be used in the future as biomarkers and serve as potential therapeutic targets in EAC.

Seom guidelines for the treatment of gastric cancer 2015

Gastric cancer is the fourth cause of death by cancer in Spain and a significant medical problem. Molecular biology results evidence that gastroesophageal junction tumors and gastric cancer should be considered as two independent entities with a different prognosis and treatment approach. Endoscopic resection in very early tumors is feasible. Neoadjuvant and adjuvant therapy in locally advanced resectable tumor increase overall survival and should be considered standard treatments. In stage IV tumors, platinum-fluoropyrimidine-based schedule, with trastuzumab in HER2-overexpressed tumors, is the firstline treatment. Different therapies in second line have demonstrated in randomized studies their clear benefit in survival improvement (AU)

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Enfermedades esofágicas: enfermedad por reflujo gastroesofágico, Barrett, acalasia y esofagitis eosinofílica / Esophageal diseases: GERD, Barrett, achalasia and eosinophilic esophagitis

En la Digestive Disease Week 2014 se han presentado importantes novedades en patología esofágica. A destacar, respecto de la enfermedad por reflujo gastroesofágico, la utilidad de la impedanciometría para el diagnóstico de la enfermedad por reflujo, o la eficacia de los inhibidores de la bomba de protones para el tratamiento del dolor torácico no coronario. Respecto del esófago de Barrett, que su prevalencia es idéntica en pacientes con y sin síntomas de reflujo, que el < 1 cm probablemente no precisa seguimiento y que en pacientes de edad y con Barrett largo, la endoscopia inicial pasa por alto hasta un 2% de lesiones significativas. Respecto de la acalasia, la miotomía quirúrgica no es superior a la dilatación endoscópica y podría ser menos efectiva que la miotomía endoscópica peroral (POEM). Respecto de la esofagitis eosinofílica, es importante tomar biopsias sistemáticamente en pacientes con disfagia, para no pasar por alto casos de esofagitis eosinofílica y que, en esta patología, la dilatación endoscópica rutinaria no solamente no parece útil para mejorar el curso de la enfermedad, sino que incluso podría empeorar la respuesta al tratamiento médico

At Digestive Disease Week (DDW) 2014, developments in esophageal disease were presented. Highlights include: the usefulness of impedancemetry to diagnose reflux disease, or the effectiveness of PPIs for treating non-cardiac chest pain. Concerning Barrett's esophagus, its prevalence is identical in patients with and without reflux symptoms, Barrett segments less than 1cm probably do not require follow-up, and in older patients with long-segment Barrett, initial endoscopies overlooked up to 2% of significant lesions. Regarding achalasia, surgical myotomy is no more effective than endoscopic dilation and may even be less effective than peroral endoscopic myotomy (POEM). In terms of eosinophilic esophagitis, it is important to systematically take biopsies in patients with dysphagia so that cases of eosinophilic esophagitis are not overlooked. In addition, for this condition, routine endoscopic dilations not only do not seem useful in improving the course of the disease, but could also worsen the response to medical treatment

Immunoreactivity of cytokeratins 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus / Imunoreatividade das citoqueratinas 7 e 20 nas células caliciformes e células colunares azuis em pacientes com evidência endoscópica de esôfago de Barrett

CONTEXT: Barrett's esophagus is characterized by the presence of goblet cells. However, when alcian-blue is utilized, another type of cells, called columnar blue cells, is frequently present in the distal esophagus of patients with endoscopic evidence of Barrett's esophagus. Cytokeratin 7 and 20 immunoreactivity has been previously studied in areas of intestinal metaplasia at the esophagogastric junction. However, the expression of these cytokeratins in columnar blue cells has not been characterized. OBJECTIVE: To compare the expression of cytokeratin 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus. METHODS: Biopsies from 86 patients with endoscopic evidence of Barrett's esophagus were evaluated. The biopsies were stained for cytokeratin 7 and 20. RESULTS: Goblet cells were present in 75 cases and columnar blue cells in 50 cases. Overall, cytokeratin 7 expression was similar in goblet cells and columnar blue cells (P = 0.25), while cytokeratin 20 was more common in goblet cells (P <0.001). In individuals with both cell types, however, cytokeratin 7 staining was the same in goblet and columnar blue cells in 95 percent of the cases, and cytokeratin 20 staining was the same in 77 percent. CONCLUSION: Goblet cells and columnar blue cells have similar immunohistochemical staining patterns for cytokeratins 7 and 20 in patients with endoscopic evidence of Barrett's esophagus.

CONTEXTO: Esôfago de Barrett é caracterizado pela presença de células caliciformes. Entretanto, quando "alcian blue" é utilizado, outro tipo de células, chamadas células colunares azuis, estão frequentemente presentes no esôfago distal de pacientes com evidência endoscópica de esôfago de Barrett. A imunoreatividade das citoqueratinas 7 e 20 tem sido estudada previamente em áreas de metaplasia intestinal na junção esôfago-gástrica. Entretanto, a expressão destas citoqueratinas nas células colunares azuis não foi caracterizada. OBJETIVO: Comparar a expressão das citoqueratinas 7 e 20 nas células caliciformes e células colunares azuis em pacientes com evidência endoscópica de esôfago de Barrett. MÉTODOS: Biopsias de 86 pacientes com evidência endoscópica de esôfago de Barrett foram avaliadas. Estas foram coradas com citoqueratinas 7 e 20. RESULTADOS: Células caliciformes estavam presentes em 75 casos e células colunares azuis em 50 casos. Ao todo, a expressão da citoqueratina 7 foi similar nas células caliciformes e células colunares azuis (P = 0,25), enquanto que a da citoqueratina 20 foi mais comum nas células caliciformes (P<0,001). Por outro lado, em indivíduos apresentando ambos os tipos de células, a coloração da citoqueratina 7 foi a mesma nas células caliciformes e células colunares azuis em 95 por cento dos casos, e a coloração da citoqueratina 20 foi a mesma em 77 por cento. CONCLUSÃO: As células caliciformes e as células colunares azuis têm padrões similares de coloração imunoistoquímica para citoqueratina 7 e 20 em pacientes com evidência endoscópica de esôfago de Barrett.

Expresión de la proteína p53 desde displasia leve hasta adenocarcinoma en un paciente con esófago de Barrett: un estudio con inmunohistoquimia / p53 protein expresion from mild dysplasia to adenocarcinoma in a patient with Barrett s esophagus: an immunohistochemical study

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O critério de positividade para a análise imunoistoquímica da p53 na confirmação da displasia do esôfago de Barrett faz diferença? / Does positive criterium for p53 immunohistochemical analysis in the confirmation of Barrett's dysplasia make difference?

RACIONAL: O esôfago de Barrett é uma complicação da doença do refluxo gastroesofágico com importante potencial de malignização. Relata-se que a expressão do marcador tumoral p53 se acentua com a progressão displasia-adenocarcinoma. OBJETIVO: Avaliar a expressão da p53 no epitélio de Barrett com presença ou não de displasia conforme dois critérios de positividade. MATERIAL E MÉTODOS: O material foi constituído por biopsias endoscópicas de 42 doentes com esôfago de Barrett. Cortes histológicos foram corados pela hematoxilina-eosina, pelo PAS-alcian blue e avaliados quanto à expressão imunoistoquímica da p53. O diagnóstico de displasia foi firmado pela concordância entre três patologistas. Foram utilizados dois critérios de positividade para a p53: 1. a coloração de, pelo menos, metade dos núcleos e 2. o encontro de qualquer núcleo corado. RESULTADOS: O número total de fragmentos foi de 229, com média de 5,4 por paciente. A displasia foi detectada em seis (14,3 por cento) casos. Para diferentes critérios de positividade, a p53 foi detectada, respectivamente, em 5 (13,9 por cento) e 14 (38,9 por cento) com epitélio metaplásico não-displásico. Especificamente nos seis casos displásicos, a p53 foi detectada, conforme o critério de positividade, em um (16,7 por cento) e quatro (66,7 por cento) casos, respectivamente. CONCLUSÕES: Nesta pequena série, a expressão imunoistoquímica da p53, independente do critério de positividade, não foi de auxílio para a confirmação de alterações displásicas no esôfago de Barrett.

Origin of adenocarcinoma in Barrett's esophagus: P53 and Ki67 expression and histopathologic background

O esôfago de Barrett é definido como a substituição do epitélio escamoso do esôfago distal por epitélio colunar. A metaplasia intestinal no esôfago de Barrett é considerada por muitos como o principal fator de risco para o desenvolvimento do adenocarcinoma. Embora já descrito, o adenocarcinoma do tipo difuso e o esôfago de Barrett sem metaplasia intestinal, são raros e pouco estudados. OBJETIVO E MÉTODO: O presente estudo objetivou o cálculo da prevalência do adenocarcinoma no esôfago de Barrett, assim como a análise macroscópica e microscópica detalhada de treze pacientes operados no período de 1990 a 2002, com realização de estudo imunohistoquímico do p53 e Ki67, correlacionando o tipo de tumor com o epitélio adjacente a este. RESULTADOS: Obtivemos uma prevalência de 5,7% de adenocarcinoma em pacientes internados para tratamento cirúrgico de esôfago de Barrett . Encontraram-se tumores relativamente grandes, com média de 4,67 ± 2,28 cm, e sempre em esôfago de Barrett longo, com média de 7,71 ± 1,5 cm. Observou-se tendência de os tumores se localizarem próximos à transição escamo-colunar. O estudo histológico mostrou dois pacientes (15,4%) que apresentavam esôfago de Barrett adjacente ao tumor do tipo juncional sem presença de metaplasia intestinal. Classificaram-se os tumores segundo a classificação japonesa de Nakamura (23% de padrão diferenciado ou intestinal e 77% de padrão indiferenciado ou gástico) e pela classificação de Laurén (61% intestinais e 39% difusos). A diferença decorre da migração dos tumores microtubulares e foveolares do padrão gástrico para o tipo intestinal de Laurén. O estudo do Ki67 foi fortemente positivo em todos os pacientes, mostrando o alto índice de proliferação celular no epitélio colunar e no tumor. O p53 mostrou-se negativo em 66,7% dos pacientes no epitélio colunar e 41,7% no tumor, não mostrando correlação entre os dois materiais. CONCLUSAO: O adenocarcinoma se desenvolve sobre o esôfago de Barrett a partir do epitélio colunar misto, intestinal, bem como do juncional, apresentando padrão tanto gástrico como intestinal; portanto tumores podem se desenvolver em epitélio colunar sem metaplasia intestinal o qual também deve ser seguido, principalmente quando for extenso.