RESUMEN
Intestinal metaplasia in the esophagus (Barrett's esophagus IM, or BE-IM) and stomach (GIM) are considered precursors for esophageal and gastric adenocarcinoma, respectively. We hypothesize that BE-IM and GIM follow parallel developmental trajectories in response to differing inflammatory insults. Here, we construct a single-cell RNA-sequencing atlas, supported by protein expression studies, of the entire gastrointestinal tract spanning physiologically normal and pathologic states including gastric metaplasia in the esophagus (E-GM), BE-IM, atrophic gastritis, and GIM. We demonstrate that BE-IM and GIM share molecular features, and individual cells simultaneously possess transcriptional properties of gastric and intestinal epithelia, suggesting phenotypic mosaicism. Transcriptionally E-GM resembles atrophic gastritis; genetically, it is clonal and has a lower mutational burden than BE-IM. Finally, we show that GIM and BE-IM acquire a protumorigenic, activated fibroblast microenvironment. These findings suggest that BE-IM and GIM can be considered molecularly similar entities in adjacent organs, opening the path for shared detection and treatment strategies. SIGNIFICANCE: Our data capture the gradual molecular and phenotypic transition from a gastric to intestinal phenotype (IM) in the esophagus and stomach. Because BE-IM and GIM can predispose to cancer, this new understanding of a common developmental trajectory could pave the way for a more unified approach to detection and treatment. See related commentary by Stachler, p. 1291. This article is highlighted in the In This Issue feature, p. 1275.
Asunto(s)
Esófago de Barrett , Gastritis Atrófica , Humanos , ARN , Metaplasia/genética , Esófago/metabolismo , Esófago/patología , Esófago de Barrett/genética , Esófago de Barrett/metabolismo , Esófago de Barrett/patología , Análisis de Secuencia de ARN , Microambiente TumoralRESUMEN
BACKGROUND: Non-endoscopic cell collection devices combined with biomarkers can detect Barrett's intestinal metaplasia and early oesophageal cancer. However, assays performed on multi-cellular samples lose information about the cell source of the biomarker signal. This cross-sectional study examines whether a bespoke artificial intelligence-based computational pathology tool could ascertain the cellular origin of microRNA biomarkers, to inform interpretation of the disease pathology, and confirm biomarker validity. METHODS: The microRNA expression profiles of 110 targets were assessed with a custom multiplexed panel in a cohort of 117 individuals with reflux that took a Cytosponge test. A computational pathology tool quantified the amount of columnar epithelium present in pathology slides, and results were correlated with microRNA signals. An independent cohort of 139 Cytosponges, each from an individual patient, was used to validate the findings via qPCR. FINDINGS: Seventeen microRNAs are upregulated in BE compared to healthy squamous epithelia, of which 13 remain upregulated in dysplasia. A pathway enrichment analysis confirmed association to neoplastic and cell cycle regulation processes. Ten microRNAs positively correlated with columnar epithelium content, with miRNA-192-5p and -194-5p accurately detecting the presence of gastric cells (AUC 0.97 and 0.95). In contrast, miR-196a-5p is confirmed as a specific BE marker. INTERPRETATION: Computational pathology tools aid accurate cellular attribution of molecular signals. This innovative design with multiplex microRNA coupled with artificial intelligence has led to discovery of a quality control metric suitable for large scale application of the Cytosponge. Similar approaches could aid optimal interpretation of biomarkers for clinical use. FUNDING: Funded by the NIHR Cambridge Biomedical Research Centre, the Medical Research Council, the Rosetrees and Stoneygate Trusts, and CRUK core grants.
Asunto(s)
Esófago de Barrett , Neoplasias Esofágicas , MicroARNs , Inteligencia Artificial , Esófago de Barrett/genética , Biomarcadores/metabolismo , Estudios Transversales , Neoplasias Esofágicas/diagnóstico , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Humanos , MicroARNs/genéticaRESUMEN
We present the largest series of diffuse large B-cell lymphoma (DLBCL) in patients younger than 18 years analysed to date by gene expression profiling using Nanostring technology to identify molecular subtypes and fluorescent in situ hybridization for translocations of MYC. We show that the activated B cell-like subtype of DLBCL is exceedingly rare in children and - in contrast to adults- not associated with outcome. Furthermore, we review the current literature and demonstrate that MYC translocations are not more frequent in paediatric compared to adult DLBCL. A prognostic role of MYC in the paediatric age groups seems unlikely.
Asunto(s)
Evolución Clonal/genética , Expresión Génica , Linfoma de Células B Grandes Difuso/diagnóstico , Linfoma de Células B Grandes Difuso/genética , Proteínas Proto-Oncogénicas c-myc/genética , Translocación Genética , Adolescente , Biomarcadores de Tumor , Niño , Preescolar , Estudios de Cohortes , Femenino , Humanos , Masculino , RecurrenciaAsunto(s)
Proteínas Inhibidoras de la Diferenciación , Linfoma de Células B , Mutación , Proteínas de Neoplasias , Femenino , Humanos , Inmunohistoquímica , Proteínas Inhibidoras de la Diferenciación/genética , Proteínas Inhibidoras de la Diferenciación/metabolismo , Linfoma de Células B/genética , Linfoma de Células B/metabolismo , Linfoma de Células B/patología , Masculino , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismoRESUMEN
The differential diagnosis between Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) can be challenging. BL has been reported to express less BCL2 than DLBCL, but this issue has not been analysed systematically. BL expressing BCL2 can be considered to be MYC/BCL2 co-expressors, a feature that is associated with poorer outcome in DLBCL but that has not been correlated with outcome in BL so far. We analysed the expression of BCL2 in 150 cases of conventionally diagnosed BL using two different BCL2 antibodies. BCL2 expression was detected in 23% of the cases, though the expression varied in intensity and number of positive cells. We did not detect any relevant differences in clinical presentation and outcome between BCL2-positive and BCL2-negative BL in a subgroup of 43 cases for which detailed clinical data were available. An independent cohort of 17 BL with expression of BCL2 were analysed molecularly, with 13 of 17 cases classified as molecularly defined BL (Burkitt Lymphoma) using gene expression profiling on formalin-fixed paraffin-embedded tissues. The four lymphomas diagnosed molecularly as intermediates did not differ in clinical presentation and outcome from molecularly defined BL.
Asunto(s)
Linfoma de Burkitt/diagnóstico , Regulación Neoplásica de la Expresión Génica , Genes bcl-2 , Proteínas de Neoplasias/biosíntesis , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Adolescente , Linfoma de Burkitt/genética , Linfoma de Burkitt/patología , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Formaldehído , Genes myc , Humanos , Linfoma de Células B Grandes Difuso/diagnóstico , Masculino , Proteínas de Neoplasias/genética , Adhesión en Parafina , Fijación del Tejido , Translocación Genética , Resultado del Tratamiento , Adulto JovenRESUMEN
Mantle cell lymphoma (MCL) is characterized by the translocation t(11;14)(q13;q32) leading to an overexpression of cyclin D1, a mediator of G1-S phase transition. Thus MCL is regarded as a paradigm of lymphoma with a dysregulated cell cycle. The proliferation rate of MCL is in fact a strong predictor of outcome. We analyzed proteins that are expressed at defined cell cycle phases, such as Ki67, survivin and phosphorylated histone H3 as well as cyclin D1, p53 and p27, on the cellular level by immunofluorescence double stainings in MCL biopsy specimens. Unexpectedly, we did not detect a shortening of early phases in MCL in vivo. Despite the control of the immunoglobulin enhancer, cyclin D1 was expressed in a cell cycle-dependent manner. However, the proliferating Ki67-positive tumor cells expressed low amounts of cyclin D1. Therefore, the expression of cyclin D1 appears not to be the driving factor behind the total proliferation rate of MCL.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Ciclo Celular/fisiología , Ciclina D1/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Proteínas Inhibidoras de la Apoptosis/metabolismo , Antígeno Ki-67/metabolismo , Linfoma de Células del Manto/patología , Proteína p53 Supresora de Tumor/metabolismo , Biomarcadores de Tumor/genética , Ciclina D1/genética , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Técnica del Anticuerpo Fluorescente , Humanos , Proteínas Inhibidoras de la Apoptosis/genética , Antígeno Ki-67/genética , Linfoma de Células del Manto/genética , Linfoma de Células del Manto/metabolismo , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Survivin , Proteína p53 Supresora de Tumor/genéticaAsunto(s)
Linfocitos B/metabolismo , Perfilación de la Expresión Génica/métodos , Regulación Neoplásica de la Expresión Génica , Linfoma de Células B/genética , Linfocitos B/patología , Biopsia , Formaldehído , Perfilación de la Expresión Génica/instrumentación , Humanos , Modelos Lineales , Linfoma de Células B/clasificación , Linfoma de Células B/patología , Adhesión en Parafina , Reproducibilidad de los Resultados , Fijación del TejidoRESUMEN
Nodular lymphocyte-predominant Hodgkin lymphoma and classical Hodgkin lymphoma are considered 2 distinct entities whose co-occurrence in 1 patient is extremely rare. We report a case of nodular lymphocyte-predominant Hodgkin lymphoma and classical Hodgkin lymphoma concurrently affecting the same lymph nodes in a 48-year-old male patient. Amplification and sequencing of the rearranged immunoglobulin heavy chain genes in tumor cells isolated by laser-assisted microdissection revealed identical variable, diverse and joining segment rearrangements and somatic hypermutation events, demonstrating a clonal relationship between the 2 lymphomas. The Epstein-Barr virus-encoded RNA and latent membrane protein 1 were present in the Hodgkin/Reed-Sternberg cells of the classical Hodgkin lymphoma but not in the tumor cells of the nodular lymphocyte-predominant Hodgkin lymphoma, pointing to a common precursor cell but differences in the early steps of pathogenesis.
Asunto(s)
Linfoma Compuesto/patología , Infecciones por Virus de Epstein-Barr/patología , Herpesvirus Humano 4/aislamiento & purificación , Enfermedad de Hodgkin/patología , Linfocitos/patología , Células Clonales , Linfoma Compuesto/inmunología , Linfoma Compuesto/virología , Infecciones por Virus de Epstein-Barr/complicaciones , Infecciones por Virus de Epstein-Barr/inmunología , Herpesvirus Humano 4/genética , Enfermedad de Hodgkin/inmunología , Enfermedad de Hodgkin/virología , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Captura por Microdisección con Láser , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/patología , Linfocitos/virología , Masculino , Persona de Mediana Edad , ARN Viral/aislamiento & purificación , Células de Reed-Sternberg/inmunología , Células de Reed-Sternberg/patología , Células de Reed-Sternberg/virología , Proteínas de la Matriz ViralRESUMEN
The V600E mutation of the BRAF gene has been reported to be associated with poor prognosis of germ cell tumors in adult patients. We analyzed the mutational status of the BRAF and KRAS gene as well as MLH1 and MSH6 expression as surrogate markers for microsatellite instability in 70 pediatric germ cell tumors. Neither BRAF and KRAS mutations nor loss of MLH1 and MSH6 expression were found. Our data provide further evidence for patient age related biological differences in germ cell tumors and demonstrate that prognostic biomarkers cannot necessarily be transferred from one age group to the other.