Vitamin D and Wnt3A have additive and partially overlapping modulatory effects on gene expression and phenotype in human colon fibroblasts

The Wnt/β-catenin signalling pathway is essential for intestinal epithelium homeostasis, but its aberrant activation is a hallmark of colorectal cancer (CRC). Several studies indicate that the bioactive vitamin D metabolite 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) inhibits proliferation and promotes epithelial differentiation of colon carcinoma cells in part through antagonism of the Wnt/β-catenin pathway. It is now accepted that stromal fibroblasts are crucial in healthy and pathologic intestine: pericryptal myofibroblasts are constituents of the stem cell niche and cancer-associated fibroblasts (CAFs) contribute to CRC progression. However, studies on the combined action of 1,25(OH)2D3 and Wnt factors in colon fibroblasts are lacking. Here we show by global transcriptomic studies that 1,25(OH)2D3 and Wnt3A have profound, additive, partially overlapping effects on the gene expression profile of CCD-18Co human colon myofibroblasts. Moreover, 1,25(OH)2D3 and Wnt3A inhibit CCD-18Co cell proliferation and migration, while 1,25(OH)2D3 reduces, but Wnt3A increases, their capacity to contract collagen gels (a marker of fibroblast activation). These data were largely confirmed in patient-derived primary colon normal fibroblasts and CAFs, and in fibroblasts from other origins. Our results indicate that 1,25(OH)2D3 and Wnt3A are strong regulators of colon fibroblast biology and contribute to a better knowledge of intestinal homeostasis and stromal fibroblast action in CRCThe work in the authors’ laboratories is supported by the Spanish Ministerio de Ciencia, Innovación y Universidades - Fondo Europeo de Desarrollo Regional (FEDER) (SAF2016-76377-R, SAF2017-90604-REDT), Consejo Superior de Investigaciones Científicas (201820I058), and Instituto de Salud Carlos III - FEDER (CIBERONC, CB16/12/00273; CIBERES, CB15/00037

Prospective Exploratory Analysis of Angiogenic Biomarkers in Peripheral Blood in Advanced NSCLC Patients Treated With Bevacizumab Plus Chemotherapy: The ANGIOMET Study

Finding angiogenic prognostic markers in advanced non-small-cell lung cancer is still an unmet medical need. We explored a set of genetic variants in the VEGF-pathway as potential biomarkers to predict clinical outcomes of patients with non-small-cell lung cancer treated with chemotherapy plus bevacizumab. We prospectively analyzed the relationship between VEGF-pathway components with both pathological and prognostic variables in response to chemotherapy plus bevacizumab in 168 patients with non-squamous non-small-cell lung cancer. Circulating levels of VEGF and VEGFR2 and expression of specific endothelial surface markers and single-nucleotide polymorphisms in VEGF-pathway genes were analyzed. The primary clinical endpoint was progression-free survival. Secondary endpoints included overall survival and objective tumor response. VEGFR-1 rs9582036 variants AA/AC were associated with increased progression-free survival (p = 0.012 and p = 0.035, respectively), and with improved overall survival (p = 0.019) with respect to CC allele. Patients with VEGF-A rs3025039 harboring allele TT had also reduced mortality risk (p = 0.049) compared with the CC allele. The VEGF-A rs833061 variant was found to be related with response to treatment, with 61.1% of patients harboring the CC allele achieving partial treatment response. High pre-treatment circulating levels of VEGF-A were associated with shorter progression-free survival (p = 0.036). In conclusion, in this prospective study, genetic variants in VEGFR-1 and VEGF-A and plasma levels of VEGF-A were associated with clinical benefit, progression-free survival, or overall survival in a cohort of advanced non-squamous non-small-cell lung cancer patients receiving chemotherapy plus antiangiogenic therapy

Assessment of a New ROS1 Immunohistochemistry Clone (SP384) for the Identification of ROS1 Rearrangements in Patients with Non–Small Cell Lung Carcinoma: the ROSING Study

Introduction: The ROS1 gene rearrangement has become an important biomarker in NSCLC. The College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology testing guidelines support the use of ROS1 immunohistochemistry (IHC) as a screening test, followed by confirmation with fluorescence in situ hybridization (FISH) or a molecular test in all positive results. We have evaluated a novel anti-ROS1 IHC antibody (SP384) in a large multicenter series to obtain real-world data. Methods: A total of 43 ROS1 FISH-positive and 193 ROS1 FISH-negative NSCLC samples were studied. All specimens were screened by using two antibodies (clone D4D6 from Cell Signaling Technology and clone SP384 from Ventana Medical Systems), and the different interpretation criteria were compared with break-apart FISH (Vysis). FISH-positive samples were also analyzed with next-generation sequencing (Oncomine Dx Target Test Panel, Thermo Fisher Scientific). Results: An H-score of 150 or higher or the presence of at least 70% of tumor cells with an intensity of staining of 2+ or higher by the SP384 clone was the optimal cutoff value (both with 93% sensitivity and 100% specificity). The D4D6 clone showed similar results, with an H-score of at least 100 (91% sensitivity and 100% specificity). ROS1 expression in normal lung was more frequent with use of the SP384 clone (p < 0.0001). The ezrin gene (EZR)-ROS1 variant was associated with membranous staining and an isolated green signal FISH pattern (p = 0.001 and p = 0.017, respectively). Conclusions: The new SP384 ROS1 IHC clone showed excellent sensitivity without compromising specificity, so it is another excellent analytical option for the proposed testing algorithm

La Inteligencia Creativa en el Proyecto de Arquitectura_PID_17_18_Anexos 1 y 2

Innovación EducativaEl PID desarrollado se sustancia en la puesta en valor del paradigma de la Inteligencia Emocional a través de un instrumento de aplicación al mecanismo enseñanza-aprendizaje de los Proyectos Arquitectónicos. Para ello se ha propuesto a los alumnos el reconocimiento y aplicación de los Sistemas de Representación Neurolingüística (visual, auditiva, y cinestésica), como metodología. Es decir, entender el Proyecto de arquitectura no como la producción de objeto, sino como instrumento de comunicación de su idea arquitectónica. Cada alumno debe reconocer, en primer lugar, la manera en que percibe el mudo (visual, auditiva, y cinestésica), de manera que esta forma de entender y comprender se convierta en activo de su propio Proyecto. El alumno no solo aplicará los conocimientos adquiridos, sino que debe incorporar de forma activa su propia manera de entender y percibir. Con este fundamento el Proyecto se inicia con la investigación de arquitecturas de programa funcional e interpretación del lugar, próximas al objeto de desarrollo de su Proyecto. En esta parte el estudiante debe reencontrase con su propia manera de expresar arquitectonicamente el edificio estudiado. Este sistema de representación de la arquitectura será el que debe sostener, desde su consciencia, en la manera de trasmitir la idea de su Proyecto.Teoría de la Arquitectura y Proyectos Arquitectónico

Memoria Final PID-17-18_060

Innovación EducativaEl PID 060 desarrollado se sustancia en la puesta en valor del paradigma de la Inteligencia Emocional a través de un instrumento de aplicación al mecanismo enseñanza-aprendizaje de los Proyectos Arquitectónicos. Para ello se ha propuesto a los alumnos el reconocimiento y aplicación de los Sistemas de Representación Neurolingüística (visual, auditiva, y cinestésica), como metodología. Es decir, entender el Proyecto de arquitectura no como la producción de objeto, sino como instrumento de comunicación de su idea arquitectónica. Cada alumno debe reconocer, en primer lugar, la manera en que percibe el mudo (visual, auditiva, y cinestésica), de manera que esta forma de entender y comprender se convierta en activo de su propio Proyecto. El alumno no solo aplicará los conocimientos adquiridos, sino que debe incorporar de forma activa su propia manera de entender y percibir. Con este fundamento el Proyecto se inicia con la investigación de arquitecturas de programa funcional e interpretación del lugar, próximas al objeto de desarrollo de su Proyecto. En esta parte el estudiante debe reencontrase con su propia manera de expresar arquitectonicamente el edificio estudiado. Este sistema de representación de la arquitectura será el que debe sostener, desde su consciencia, en la manera de trasmitir la idea de su Proyecto

Clinical management of epidermal growth factor receptor mutation-positive non-small cell lung cancer patients after progression on previous epidermal growth factor receptor tyrosine kinase inhibitors : The necessity of repeated molecular analysis

One of the most important advances in the treatment of non-small cell lung cancer (NSCLC) has been the identification of molecular alterations vulnerable to targeted inhibition, such as mutations in the epidermal growth factor receptor (EGFR) gene. EGFR tyrosine kinase inhibitors (EGFR-TKIs) are targeted agents used to treat EGFR mutation-positive advanced NSCLC showing significant improvements in terms of response rate (RR) and progression-free survival (PFS) compared to conventional chemotherapy. However, all patients eventually develop resistance to first-line EGFR-TKIs. The most common mechanism of acquired resistance is the secondary acquisition of a single missense mutation within exon 20 in the EGFR gene, known as the T790M mutation (49-60%). New agents targeting the T790M mutation have undergone clinical development, and among these, osimertinib has shown significant activity in relapsing EGFR mutation positive patients harbouring the T790M mutation. Although precision medicine is a reality for NSCLC, obtaining relevant tissue for repeated molecular analysis from these patients remains a challenge. In this article, a group of experts from the Spanish Society of Medical Oncology (SEOM) and the Spanish Lung Cancer Group (GECP) evaluated the role of rebiopsy and the potential application of plasma-testing methodologies in advanced EGFR mutation patients progressing after EGFR-TKI

Gene regulatory and phenotypic effects of calcitriol and canonical WNT in human colon fibroblasts

Trabajo presentado en el 42nd Congress of the Spanish Society of Biochemistry and Molecular Biology, celebrado en Madrid (España), del 16 al 19 de julio de 2019.Vitamin D3 (cholecalciferol) is synthesized in the skin by action of solar UV radiation or incorporated via diet, and is the inactive precursor of 1α,25-dihydroxyvitamin D3 (calcitriol), a major pleiotropic hormone in the human organism. By binding and activation of vitamin D receptor (VDR), a member of the superfamily of nuclear receptors, calcitriol regulates the expression of hundreds of genes in a tissue- and cell-type specific manner by transcriptional and posttranscriptional mechanisms. Human WNT factors are a family of 19 members of secreted proteins that regulate crucial processes in many tissues and organs during development and adult life. Some (canonical) WNTs act by modulating the transcriptional activity of β-catenin whereas other (non-canonical) WNTs affect different signal transducers (Ca2+, Rho, JNK...) independently of β-catenin. Colorectal cancer (CRC) is one of the most important neoplasias worldwide in terms of incidence and mortality. The key role of the constitutive activation of the WNT/β-catenin signaling pathwaypromoting CRC and the protective effect of VDR agonists, in part by antagonizing the WNT/β-catenin pathway, have been widely described. However, the effects of calcitriol and canonical WNTs on stromal fibroblasts, which are important players in CRC with protumorigenic action, remain mostly unknown. We have studied the effect of calcitriol and WNT3A on the human CCD-18Co colonic fibroblast cell line and on primary colon fibroblasts isolated from CRC patients. Data from global transcriptomic analyses by RNA-sequencing and also functional assays (cell proliferation, migration...) indicate that calcitriol and WNT3A exert a wide regulatory action on the gene expression and phenotype of colonic fibroblasts. Remarkably, a complex interplay exists between the two agents: while calcitriol and WNT3A act cooperatively in some effects, antagonistic actions are found in others. Our results show that calcitriol and WNT3A are important modulators of human colon fibroblasts, with potential implications in colon homeostasis and neoplastic transformation

Gene regulatory and phenotypic effects of calcitriol and canonical WNT in human colon fibroblasts

Trabajo presentado en el 6th Symposium on Biomedical Research >Advances and Perspectives in Molecular Endocrinology>, celebrado en Madrid (España) el 31 de mayo de 2019.Vitamin D3 (cholecalciferol) is synthesized in the skin by action of solar UV radiation or incorporated via diet, and is the inactive precursor of 1α,25-dihydroxyvitamin D3 (calcitriol), a major pleiotropic hormone in the human organism. By binding and activation of vitamin D receptor (VDR), a member of the superfamily of nuclear receptors, calcitriol regulates the expression of hundreds of genes in a tissue- and cell-type specific manner by transcriptional and posttranscriptional mechanisms. Human WNT factors are a family of 19 members of secreted proteins that regulate crucial processes in many tissues and organs during development and adult life. Some (canonical) WNTs act by modulating the transcriptional activity of β-catenin whereas other (non-canonical) WNTs affect different signal transducers (Ca2+, Rho, JNK...) independently of β-catenin. Colorectal cancer (CRC) is one of the most important neoplasias worldwide in terms of incidence and mortality. The key role of the constitutive activation of the WNT/β-catenin signaling pathway promoting CRC and the protective effect of VDR agonists, in part by antagonizing the WNT/β-catenin pathway, have been widely described. However, the effects of calcitriol and canonical WNTs on stromal fibroblasts, which are important players in CRC with protumorigenic action, remain mostly unknown. We have studied the effect of calcitriol and WNT3A on the human CCD-18Co colonic fibroblast cell line and on primary colon fibroblasts isolated from CRC patients. Data from global transcriptomic analyses by RNA-sequencing and also functional assays (cell proliferation, migration...) indicate that calcitriol and WNT3A exert a wide regulatory action on the gene expression and phenotype of colonic fibroblasts. Remarkably, a complex interplay exists between the two agents: while calcitriol and WNT3A act cooperatively in some effects, antagonistic actions are found in others. Our results show that calcitriol and WNT3A are important modulators of human colon fibroblasts, with potential implications in colon homeostasis and neoplastic transformation.Ministerio de Ciencia, Innovación y Universidades (SAF2016-76377-R, Nurcamein2) and Instituto de Salud Carlos III (CIBERONC) of Spain - Fondo Europeo de Desarrollo Regional

Milicias y tropas negras de Buenos Aires. Afro argentinos armados para defender a sus amos

The city of Buenos Aires was founded, for the second time 1580 essentially as an outpost of the Spanish Empire to be protected from Indian attacks and possible foreign incursion, basically Portuguese in origin. Thus, from the beginnings of the city's founding, African slaves were used for the defense of the city. They acted in the urban militias, and were outstanding in the defense and re-conquest of the city when the English attacked in 1806- 1807. Innumerable prizes and eulogies were followed by integration of segregated divisions with a white officer corps. After 1813, a series of decrees ordered owners to exchange slaves for use in the Wars of Independence. Thus, property owners were obliged to sell to the State a number of their slaves according to the work performed by the slaves. The slaves would enter the armies as free persons and would serve five years as 'front line" soldiers in order to obtain their freedom. This article studies the participation of the Afro-Argentines in the distinct 'fronts" of the armies of liberation. The paper also reflects on the opinions of the generals who commanded Afro-Argentine troops.La ciudad de Buenos Aires fue fundada, por segunda vez, en 1580 esencialmente como un puesto de avanzada para que los españoles se protegieran de los indígenas y de una posible incursión extranjera de origen portugués. Por lo tanto desde la fundación de la ciudad los esclavos africanos fueron empleados para su defensa. Los esclavos actuaban en milicias urbanas y sus acciones fueron decisivas para la defensa y reconquista de Buenos Aires cuando los ingleses la atacaron en 1806-1807. Innumerables premios y elogios fueron seguidos por la integración de las divisiones separadas, con un cuerpo de oficiales blancos. Después de 1813, una serie de decretos ordenaba a los dueños entregar a sus esclavos para las guerras de Independencia . De esta manera los esclavos entraron al ejército como personas libres y debían servir por cinco años en el frente para obtener su libertad definitiva. Este artículo estudia la participación de afro-argentinos en distintos frentes de los ejércitos de liberación. El ensayo también refleja las opiniones de los generales que comandaban las tropas afro-argentinas

Clinical management of epidermal growth factor receptor mutation-positive non-small cell lung cancer patients after progression on previous epidermal growth factor receptor tyrosine kinase inhibitors: the necessity of repeated molecular analysis

of the most important advances in the treatment of non-small cell lung cancer (NSCLC) has been the identification of molecular alterations vulnerable to targeted inhibition, such as mutations in the epidermal growth factor receptor (EGFR) gene. EGFR tyrosine kinase inhibitors (EGFR-TKIs) are targeted agents used to treat EGFR mutation-positive advanced NSCLC showing significant improvements in terms of response rate (RR) and progression-free survival (PFS) compared to conventional chemotherapy. However, all patients eventually develop resistance to first-line EGFR-TKIs. The most common mechanism of acquired resistance is the secondary acquisition of a single missense mutation within exon 20 in the EGFR gene, known as the T790M mutation (49-60%). New agents targeting the T790M mutation have undergone clinical development, and among these, osimertinib has shown significant activity in relapsing EGFR mutation positive patients harbouring the T790M mutation. Although precision medicine is a reality for NSCLC, obtaining relevant tissue for repeated molecular analysis from these patients remains a challenge. In this article, a group of experts from the Spanish Society of Medical Oncology (SEOM) and the Spanish Lung Cancer Group (GECP) evaluated the role of rebiopsy and the potential application of plasma-testing methodologies in advanced EGFR mutation patients progressing after EGFR-TKI