GD3 synthase overexpression sensitizes hepatocarcinoma cells to hypoxia and reduces tumor growth by suppressing the cSrc/NF-κB survival pathway

This is an open-access article distributed under the terms of the Creative Commons Attribution License.[Background]: Hypoxia-mediated HIF-1a stabilization and NF-kB activation play a key role in carcinogenesis by fostering cancer cell survival, angiogenesis and tumor invasion. Gangliosides are integral components of biological membranes with an increasingly recognized role as signaling intermediates. In particular, ganglioside GD3 has been characterized as a proapoptotic lipid effector by promoting cell death signaling and suppression of survival pathways. Thus, our aim was to analyze the role of GD3 in hypoxia susceptibility of epatocarcinoma cells and in vivo tumor growth.[Methodology/Principal Findings]: We generated and characterized a human hepatocarcinoma cell line stably expressing GD3 synthase (Hep3B-GD3), which catalyzes the synthesis of GD3 from GM3. Despite increased GD3 levels (2–3 fold), no significant changes in cell morphology or growth were observed in Hep3B-GD3 cells compared to wild type Hep3B cells under normoxia. However, exposure of Hep3B-GD3 cells to hypoxia (2% O2) enhanced reactive oxygen species (ROS) generation, resulting in decreased cell survival, with similar findings observed in Hep3B cells exposed to increasing doses of exogenous GD3. In addition, hypoxia-induced c-Src phosphorylation at tyrosine residues, NF-kB activation and subsequent expression of Mn-SOD were observed in Hep3B cells but not in Hep3B-GD3 cells. Moreover, MnTBAP, an antioxidant with predominant SOD mimetic activity, reduced ROS generation, protecting Hep3B-GD3 cells from hypoxia-induced death. Finally, lower tumor growth, higher cell death and reduced Mn-SOD expression were observed in Hep3B-GD3 compared to Hep3B tumor xenografts.[Conclusion]: These findings underscore a role for GD3 in hypoxia susceptibility by disabling the c-Src/NF-kB survival pathway resulting in lower Mn-SOD expression, which may be of relevance in hepatocellular carcinoma therapy.Grant support: CIBEREHD and grants FIS06/0395, FIS07/1039, SAF2006-06789 and SAF2008-02199 by Instituto de Salud Carlos III and Ministry of Science and Innovation from Spain, and from the Research Center for Liver and Pancreatic Diseases, P50-AA-11999 funded by the US National Institute on Alcohol Abuse and Alcoholism.Peer reviewe

Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer

Hepatocellular carcinoma (HCC) frequently develops in a pro-inflammatory and pro-fibrogenic environment with hepatic stellate cells (HSCs) remodeling the extracellular matrix composition. Molecules secreted by liver tumors contributing to HSC activation and peritumoral stromal transformation remain to be fully identified. Here we show that conditioned medium from HCC cell lines, Hep3B and HepG2, induced primary mouse HSCs transdifferentiation, characterized by profibrotic properties and collagen modification, with similar results seen in the human HSC cell line LX2. Moreover, tumor growth was enhanced by coinjection of HepG2/LX2 cells in a xenograft murine model, supporting a HCC-HSC crosstalk in liver tumor progression. Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules. In fact, recombinant angiogenin induced in vitro HSC activation requiring its nuclear translocation and rRNA transcriptional stimulation. Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin. Finally, neomycin administration reduced tumor growth of HepG2-LX2 cells coinjected in mice. In conclusion, angiogenin secretion by HCCs favors tumor development by inducing HSC activation and ECM remodeling. These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC managementThis study was funded by grants from the Instituto de Salud Carlos III (FIS PI12/00110, PI09/00056 to A.M., FIS PI10/02114, PI13/00374 to M.M., PI12/01265 to J.C. and PI11/0325 to J.F.C.), Ministerio de Economía y Competitividad (SAF 2012/34831 to J.F.C. and SAF2011-23031 to C.G.R.) and co-funded by FEDER (Fondo Europeo de Desarrollo Regional, Unión Europea. “Una manera de hacer Europa”); center grant P50-AA-11999 from Research Center for Liver and Pancreatic Diseases, US NIAAA to J.F.C.); Fundació la Marató de TV3 to J.F.C., Mutua Madrilea (AP103502012) to C.G.R., and by CIBERehd from the Instituto de Salud Carlos IIIPeer Reviewe

Genetic association study of dyslexia and ADHD candidate genes in a Spanish cohort: Implications of comorbid samples

Published: October 31, 2018Dyslexia and attention deficit hyperactivity disorder (ADHD) are two complex neuro-behaviorally disorders that co-occur more often than expected, so that reading disability has been linked to inattention symptoms. We examined 4 SNPs located on genes previously associated to dyslexia (KIAA0319, DCDC2, DYX1C1 and FOXP2) and 3 SNPs within genes related to ADHD (COMT, MAOA and DBH) in a cohort of Spanish children (N = 2078) that met the criteria of having one, both or none of these disorders (dyslexia and ADHD). We used a case-control approach comparing different groups of samples based on each individual diagnosis. In addition, we also performed a quantitative trait analysis with psychometric measures on the general population (N = 3357). The results indicated that the significance values for some markers change depending on the phenotypic groups compared and/or when considering pair-wise marker interactions. Furthermore, our quantitative trait study showed significant genetic associations with specific cognitive processes. These outcomes advocate the importance of establishing rigorous and homogeneous criteria for the diagnosis of cognitive disorders, as well as the relevance of considering cognitive endophenotypes.The work of MSM and MC was supported by CONSOLIDER-Ingenio- 2010_COEDUCA (CSD2008-00048). AMA, LB and AG-L’s work was supported by the Basque Department of Industry, Tourism and Trade (Etortek Program), Innovation Technology Department of Bizkaia and CIBERehd Network. MC was also supported by grants (PSI2015-67353-R), and Ayuda Centro de Excelencia Severo Ochoa SEV-2015-0490 from the MINECO, and by grant (ERC-2011-ADG-295362) from the European Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Low-count monoclonal B-cell lymphocytosis persists after seven years of follow up and is associated with a poorer outcome

Low-count monoclonal B-cell lymphocytosis is defined by the presence of very low numbers of circulating clonal B cells, usually phenotypically similar to chronic lymphocytic leukemia cells, whose biological and clinical significance remains elusive. Herein, we re-evaluated 65/91 low-count monoclonal B-cell lymphocytosis cases (54 chronic lymphocytic leukemia-like and 11 non-chronic lymphocytic leukemia-like) followed-up for a median of seven years, using high-sensitivity flow cytometry and interphase fluorescence in situ hybridization. Overall, the clone size significantly increased in 69% of low-count monoclonal B-cell lymphocytosis cases, but only one subject progressed to high-count monoclonal B-cell lymphocytosis. In parallel, the frequency of cytogenetic alterations increased over time (32% vs. 61% of cases, respectively). The absolute number of the major T-cell and natural killer cell populations also increased, but only among chronic lymphocytic leukemia-like cases with increased clone size vs. age- and sex-matched controls. Although progression to chronic lymphocytic leukemia was not observed, the overall survival of low-count monoclonal B-cell lymphocytosis individuals was significantly reduced vs. non-monoclonal B-cell lymphocytosis controls (P=0.03) plus the general population from the same region (P≤0.001), particularly among females (P=0.01); infection and cancer were the main causes of death in low-count monoclonal B-cell lymphocytosis. In summary, despite the fact that mid-term progression from low-count monoclonal B-cell lymphocytosis to high-count monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia appears to be unlikely, these clones persist at increased numbers, usually carrying more genetic alterations, and might thus be a marker of an impaired immune system indirectly associated with a poorer outcome, particularly among females.This work was supported by the RD06/0020/0035 and RD12/0036/0048 grants from Red Temática de Investigación Cooperativa en Cáncer (RTICC), Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, (Madrid, Spain and FONDOS FEDER); CB16/12/00400 grant (CIBERONC, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, (Madrid, Spain and FONDOS FEDER); the FIS PI06/0824-FEDER, PS09/02430-FEDER, PI12/00905- FEDER, DTS15/00119-FEDER, PI16/00787-FEDER and PI17/00399-FEDER grants, from the Fondo de Investigación Sanitaria of Instituto de Salud Carlos III; the GRS206/A/08 grant, (Ayuda al Grupo GR37 de Excelencia, SAN/1778/2009) from the Gerencia Regional de Salud (Consejería de Educación and Consejería de Sanidad of Castilla y León, Valladolid, Spain) and the SA079U14 grant (Consejería de Educación and Consejería de Sanidad of Castilla y León, Valladolid, Spain). ML Gutiérrez is supported by grant PTA2014-09963-I from the Instituto de Salud Carlos III

Cytogenetic profiles in multiple myeloma and monoclonal gammopathy of undetermined significance: a study in highly purified aberrant plasma cells

This is an open-access paper.Cytogenetic studies in clonal plasma cell disorders have mainly been done in whole bone marrow or CD138+ microbead-enriched plasma cells and suggest that recurrent immunoglobulin heavy chain translocations - e.g. t(4;14) - are primary oncogenetic events. The aim of this study was to determine cytogenetic patterns of highly purified aberrant plasma cells (median purity ≥98%) in different clonal plasma cell disorders. We analyzed aberrant plasma cells from 208 patients with multiple myeloma (n=148) and monoclonal gammopathy of undetermined significance (n=60) for the presence of del(13q14), del(17p13) and t(14q32) using multicolor interphase fluorescence in situ hybridization. Additionally, immunoglobulin heavy chain gene arrangements were analyzed and complementarity determining region 3 was sequenced in a subset of patients and combined multicolor interphase fluorescence in situ hybridization/immunofluorescent protein staining analyses were performed in selected cases to confirm clonality and cytogenetic findings. At diagnosis, 96% of cases with multiple myeloma versus 77% of monoclonal gammopathy of undetermined significance cases showed at least one cytogenetic alteration and/or hyperdiploidy. The cytogenetic heterogeneity of individual cases reflected coexistence of cytogenetically-defined aberrant plasma cell clones, and led to the assumption that karyotypic alterations were acquired stepwise. Cases of multiple myeloma and monoclonal gammopathy of undetermined significance frequently showed different but related cytogenetic profiles when other cytogenetic alterations such as deletions/gains of the immunoglobulin heavy chain or the fibroblast growth factor receptor 3 were additionally considered. Interestingly, in 24% of multiple myeloma versus 62% of monoclonal gammopathy of undetermined significance patients with an immunoglobulin heavy chain translocation, aberrant plasma cells with and without t(14q32) coexisted in the same patient. Our data suggest that recurrent immunoglobulin heavy chain translocations might be absent in the primordial plasma cell clone in a significant proportion of patients with clonal plasma cell disorders carrying these cytogenetic alterations.This work was partially supported by grants from the Fundacion Memoria de Don Samuel Solorzano Barruso, Salamanca, Spain (FS/4-2010). The authors would also like to thank the Dr. Werner Jackstädt Foundation (Wuppertal, Germany) for grant supporting the work of Martin Schmidt-Hieber. The authors would like to thank the Cooperative Research Thematic Network (RTICs; RTICC RD06/0020/0035, RD06/0020/0006 and G03/136), MM Jevitt, SL firm, Instituto de Salud Carlos III/Subdirección General de Investigación Sanitaria (FIS: PI060339; 02/0905; 01/0089/01-02; PS09/01897) and Gerencia Regional de Salud de Castilla y León; Ayuda de Excelencia de Castilla y León, Consejeria de Educación (EDU/894/2009, GR37), and Consejería de Sanidad (557/A/10), Junta de Castilla y León, Valladolid, Spain for supporting this study. JMS is supported by a grant (CP05/00321) from the ISCIII, Ministerio de Ciencia e Innovacion, Madrid, Spain.Peer Reviewe

Characterizing storm-induced coastal change hazards along the United States West Coast

Traditional methods to assess the probability of storm-induced erosion and flooding from extreme water levels have limited use along the U.S. West Coast where swell dominates erosion and storm surge is limited. This effort presents methodology to assess the probability of erosion and flooding for the U.S. West Coast from extreme total water levels (TWLs), but the approach is applicable to coastal settings worldwide. TWLs were derived from 61 years of wave and water level data at shore-perpendicular transects every 100-m along open coast shorelines. At each location, wave data from the Global Ocean Waves model were downscaled to the nearshore and used to empirically calculate wave run-up. Tides were simulated using the Oregon State University?s tidal data inversion model and non-tidal residuals were calculated from sea-surface temperature and pressure anomalies. Wave run-up was combined with still water levels to generate hourly TWL estimates and extreme TWLs for multiple return periods. Extremes were compared to onshore morphology to determine erosion hazards and define the probability of collision, overwash, and inundation

N7-methylguanosine methylation of tRNAs regulates survival to stress in cancer

Tumour progression and therapy tolerance are highly regulated and complex processes largely dependent on the plasticity of cancer cells and their capacity to respond to stress. The higher plasticity of cancer cells highlights the need for identifying targetable molecular pathways that challenge cancer cell survival. Here, we show that

Características y manejo del cáncer de mama precoz en mujeres añosas asistidas en la Unidad Docente Asistencial de Mastología del Hospital de Clínicas en el período 2011-2018

Nota de contribución: Concepción y diseño del estudio: Camejo N. y Castillo C.-- Hernández A. y Delgado N.-- Recolección de los datos o realización de los experimentos: Camila Goldman, Vanra Barcena, Santiago González, Sofia Arenas, Martina Abero y María Icardo.-- Análisis e interpretación de los datos o resultados: Camejo N, Camila Goldman, Vanra Bárcena, Santiago González, Sofia Arenas, Martina Abero y María Icardo.-- Análisis estadísticos de los datos o resultados: Camejo N., Hernández A., Camila Goldman, Vanra Bárcena, Santiago González, Sofia Arenas, Martina Abero y María Icardo.-- Elaboración de un borrador del manuscrito: Camejo N., Castillo C. y Hernández A.-- Escritura del manuscrito: Camejo N., Castillo C. y Hernández A.--Supervisión del trabajo: Delgado L.Nota del Editor: El editor responsable para la publicación de este manuscrito es Juan Dapueto.Introducción: Existen pocas pautas para el tratamiento del cáncer de mama (CM) en pacientes añosas, lo que puede conducir al sub o sobre tratamiento. Objetivo: Conocer las características, manejo y la evolución del CM precoz en mujeres añosas. Material y métodos: Estudio observacional, descriptivo, transversal. Se recolectaron datos relacionados con las características clínico patológicas y la evolución de pacientes de 70 años o más tratadas por CM en el período comprendido entre el 1/1/ 2011 y el 31/12/ 2018, asistidas en el Hospital de Clínicas. Se utilizaron herramientas de estadística descriptiva y para calcular la sobrevida global (SVG) se utilizó el método de Kaplan-Meier. Resultados: se incluyeron 31 pacientes; la edad mediana al diagnóstico fue 76,8 años; las características clínico-patológicas fueron: carcinoma ductal: 71%; GH 1-2: 74,2%; estadio I: 54,8 %; sin metástasis axilares: 80,6 %; HER2-RE/RP+ 80,6%; HER2+ 16,7%, y triple negativas 3,2%. El 29% de las pacientes fueron diagnosticadas mediante tamizaje poblacional y el 74,2% recibieron tratamiento según pautas vigentes, mientras que el 38,7% fueron subtratadas y el 16,1% sobretratadas. La mediana de SVG fue de 98,7 meses. Conclusiones: Una minoría de las pacientes fue diagnosticada mediante tamizaje oblacional, el tipo histológico más frecuente fue el ductal y la prevalencia de los tumores HER2-RE/RP+ fue mayor que en las pacientes más jóvenes. La mayoría de las pacientes recibió tratamiento estándar.Introduction: There are few guidelines for the treatment of breast cancer (BC) in older patients, which can lead to under- or over treatment. Objective: To understand the characteristics, management and evolution of early BC in older women. Material and methods: Observational, descriptive, cross-sectional study. Data were collected on the clinical-pathological characteristics and evolution of patients aged 70 years or older, treated for BC in the period from 1/1/ 2011 to 31/12/ 2018, at the Hospital de Clínicas. Descriptive statistical tools were used and the Kaplan-Meier method was applied to calculate the overall survival (OS) rate. Results: 31 patients were included; median age at diagnosis was 76.8 years old; the clinical-pathological characteristics were: ductal carcinoma: 71%; HG 1-2: 74.2%; stage I: 54.8%; no axillary metastases: 80.6%; HER2-ER/PR+ 80.6%; HER2+ 16.7%, and triple negative 3.2%. Of all the patients, 29% were diagnosed through screening and 74.2% were treated according to current guidelines, while 38.7% were under-treated and 16.1% over-treated. The median OS was 98.7 months. Conclusions: A minority of patients were diagnosed by screening, the most frequent histological type was ductal and the prevalence of HER2-RE/RP+ tumors was higher than in younger patients. Most patients received standard treatment.Introdução: Existem poucas diretrizes para o tratamento do câncer de mama (CM) em pacientes idosos, o que pode levar ao sub ou excesso de tratamento. Objetivo: Conhecer as características, manejo e evolução do MC precoce em mulheres idosas. Material e métodos: estudo observacional, descritivo e transversal. Foram coletados dados relacionados às características clínico patológicas e à evolução dos pacientes com 70 anos ou mais atendidos por CM no período de 01/01/2011 a 31/12/2018, atendidos no Hospital de Clínicas. Ferramentas de estatística descritiva foram usadas e o método de Kaplan-Meier foi usado para calcular a sobrevida global (SVG). Resultados: 31 pacientes foram incluídos; a mediana de idade ao diagnóstico foi de 76,8 anos; as características clínico-patológicas foram: carcinoma ductal: 71%; GH 1-2: 74,2%; estágio I: 54,8%; sem metástases axilares: 80,6%; HER2-RE / RP + 80,6%; HER2 + 16,7% e triplo negativo 3,2%. 29% dos pacientes foram diagnosticados por triagem populacional e 74,2% receberam tratamento de acordo com as diretrizes atuais, enquanto 38,7% foram subtratados e 16,1% supertratados. O SVG médio foi de 98,7 meses. Conclusões: A minoria dos pacientes foi diagnosticada por rastreamento populacional, o tipo histológico mais frequente foi ductal e a prevalência de tumores HER2-RE / RP + foi maior do que em pacientes mais jovens. A maioria dos pacientes recebeu tratamento padrão

Low-count monoclonal B-cell lymphocytosis persists after seven years of follow up and is associated with a poorer outcome

[EN]Low-count monoclonal B-cell lymphocytosis is defined by the presence of very low numbers of circulating clonal B cells, usually phenotypically similar to chronic lymphocytic leukemia cells, whose biological and clinical significance remains elusive. Herein, we re-evaluated 65/91 low-count monoclonal B-cell lymphocytosis cases (54 chronic lymphocytic leukemia-like and 11 non-chronic lymphocytic leukemialike) followed-up for a median of seven years, using high-sensitivity flow cytometry and interphase fluorescence in situ hybridization. Overall, the clone size significantly increased in 69% of low-count monoclonal B-cell lymphocytosis cases, but only one subject progressed to high-count monoclonal B-cell lymphocytosis. In parallel, the frequency of cytogenetic alterations increased over time (32% vs. 61% of cases, respectively). The absolute number of the major T-cell and natural killer cell populations also increased, but only among chronic lymphocytic leukemia-like cases with increased clone size vs. age- and sex-matched controls. Although progression to chronic lymphocytic leukemia was not observed, the overall survival of low-count monoclonal B-cell lymphocytosis individuals was significantly reduced vs. non-monoclonal Bcell lymphocytosis controls (P=0.03) plus the general population from the same region (P≤0.001), particularly among females (P=0.01); infection and cancer were the main causes of death in low-count monoclonal B-cell lymphocytosis. In summary, despite the fact that mid-term progression from low-count monoclonal B-cell lymphocytosis to high-count monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia appears to be unlikely, these clones persist at increased numbers, usually carrying more genetic alterations, and might thus be a marker of an impaired immune system indirectly associated with a poorer outcome, particularly among females

Phenotypic identification of subclones in multiple myeloma with different chemoresistant, cytogenetic and clonogenic potential

Knowledge about clonal diversity and selection is critical to understand multiple myeloma (MM) pathogenesis, chemoresistance and progression. If targeted therapy becomes reality, identification and monitoring of intraclonal plasma cell (PC) heterogeneity would become increasingly demanded. Here we investigated the kinetics of intraclonal heterogeneity among 116 MM patients using 23-marker multidimensional flow cytometry (MFC) and principal component analysis, at diagnosis and during minimal residual disease (MRD) monitoring. Distinct phenotypic subclones were observed in 35116 (30%) newly diagnosed MM patients. In 1035 patients, persistent MRD was detected after 9 induction cycles, and longitudinal comparison of patient-paired diagnostic vs MRD samples unraveled phenotypic clonal tiding after therapy in half (510) of the patients. After demonstrating selection of distinct phenotypic subsets by therapeutic pressure, we investigated whether distinct fluorescence-activated cell-sorted PC subclones had different clonogenic and cytogenetic profiles. In half (510) of the patients analyzed, distinct phenotypic subclones showed different clonogenic potential when co-cultured with stromal cells, and in 611 cases distinct phenotypic subclones displayed unique cytogenetic profiles by interphase fluorescence in situ hybridization, including selective del(17p13). Collectively, we unravel potential therapeutic selection of preexisting diagnostic phenotypic subclones during MRD monitoring; because phenotypically distinct PCs may show different clonogenic and cytogenetic profiles, identification and follow-up of unique phenotypic-genetic myeloma PC subclones may become relevant for tailored therapy.Peer Reviewe