Modelos preclínicos en oncología molecular: hacia una medicina personalizada

La profesora invitada ha desarrollado un taller de resolución de un caso práctico con y para los alumnos de la asignatura "Biología Molecular del Cáncer" (optativa de 4º curso del Grado en Bioquímica)El objetivo de esta actividad práctica es ilustrar algunos de los modelos preclínicos utilizados en oncología y discutir su aplicación para la implementación en la clínica de tratamientos personalizados dirigidos a dianas moleculares. Para cubrir este objetivo hemos utilizado un caso clínico figurado. Los alumnos han trabajado en grupos de 2-3 alumnos con antelación a la sesión práctica. Se les aportó información a partir de la cual se formulan una serie de preguntas para cuya contestación los alumnos debieron analizar críticamente la información aportada y establecer una estrategia de "investigación" para buscar información científica adicional necesaria para contestar apropiadamente a las preguntas. Durante el proceso de análisis e "investigación" los componentes del equipo aprenden de forma colaborativa. Cada grupo entregó un documento con sus respuestas razonadas con antelación a la fecha en que se realizó el taller de puesta en común y discusión. Esta actividad ha sido diseñada y preparada por la Profesora Dra. Benilde Jiménez Cuenca (Universidad Autónoma de Madrid), quien dirigió el taller de puesta en común y discusión el jueves 5 de mayo de 11.30 a 13.30

Keratinocyte growth factor induces angiogenesis and protects endothelial barrier function

9 pages, 6 figures, 1 table.Keratinocyte growth factor (KGF), also called fibroblast growth factor-7, is widely known as a paracrine growth and differentiation factor that is produced by mesenchymal cells and has been thought to act specifically on epithelial cells. Here it is shown to affect a new cell type, the microvascular endothelial cell. At subnanomolar concentrations KGF induced in vivo neovascularization in the rat cornea. In vitro it was not effective against endothelial cells cultured from large vessels, but did act directly on those cultured from small vessels, inducing chemotaxis with an ED50 of 0.02-0.05 ng/ml, stimulating proliferation and activating mitogen activated protein kinase (MAPK). KGF also helped to maintain the barrier function of monolayers of capillary but not aortic endothelial cells, protecting against hydrogen peroxide and vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) induced increases in permeability with an ED50 of 0.2-0.5 ng/ml. These newfound abilities of KGF to induce angiogenesis and to stabilize endothelial barriers suggest that it functions in microvascular tissue as it does in epithelial tissues to protect them against mild insults and to speed their repair after major damage.Peer reviewe

Autocrine stimulation of clear-cell renal carcinoma cell migration in hypoxia via HIF-independent suppression of thrombospondin-1

Thrombospondin-1 is a matricellular protein with potent antitumour activities, the levels of which determine the fate of many different tumours, including renal carcinomas. However, the factors that regulate this protein remain unclear. In renal carcinomas, hypoxic conditions enhance the expression of angiogenic factors that help adapt tumour cells to their hostile environment. Therefore, we hypothesized that anti-angiogenic factors should correspondingly be dampened. Indeed, we found that hypoxia decreased the thrombospondin-1 protein in several clear cell renal carcinoma cell lines (ccRCC), although no transcriptional regulation was observed. Furthermore, we proved that hypoxia stimulates multiple signals that independently contribute to diminish thrombospondin-1 in ccRCC, which include a decrease in the activity of oxygen-dependent prolylhydroxylases (PHDs) and activation of the PI3K/Akt signalling pathway. In addition, thrombospondin-1 regulation in hypoxia proved to be important for ccRCC cell migration and invasionThis work was supported by the Ministerio de Ciencia e Innovacion (MCINN) SAF2009-11113 and METOXIA, a Collaborative Project under the 7th Research Framework Programme of the European Union FP7-HEALTH-2007B (ref. HEALTH-F2-2009-222741

Identification of non-coding genetic variants in samples from hypoxemic respiratory disease patients that affect the transcriptional response to hypoxia

A wide range of diseases course with an unbalance between the consumption of oxygen by tissues and its supply. This situation triggers a transcriptional response, mediated by the hypoxia inducible factors (HIFs), that aims to restore oxygen homeostasis. Little is known about the inter-individual variation in this response and its role in the progression of disease. Herein, we sought to identify common genetic variants mapping to hypoxia response elements (HREs) and characterize their effect on transcription. To this end, we constructed a list of genome-wide HIF-binding regions from publicly available experimental datasets and studied the genetic variability in these regions by targeted re-sequencing of genomic samples from 96 chronic obstructive pulmonary disease and 144 obstructive sleep apnea patients. This study identified 14 frequent variants disrupting potential HREs. The analysis of the genomic regions containing these variants by means of reporter assays revealed that variants rs1009329, rs6593210 and rs150921338 impaired the transcriptional response to hypoxia. Finally, using genome editing we confirmed the functional role of rs6593210 in the transcriptional regulation of EGFR. In summary, we found that inter-individual variability in non-coding regions affect the response to hypoxia and could potentially impact on the progression of pulmonary diseases.Ministerio de Ciencia e Innovación (Spanish Ministry of Science and Innovation, MICINN) [SAF2011 24225 to LdelP, SAF2014-53819-R to L.delP., B.J.]; Comunidad Autónoma de Madrid [S2010/BMD-2542 to L.delP., F.G.R., J.A.], Sociedad Española de Neumología y Cirugía Torácica (SEPAR) [34/2013 to LdelP, F.G.R.]; Fondo de Investigación Sanitaria/Instituto de Salud Carlos III [PI13-01512 to F.G.R.]; Fundación Caja Madrid (Beca de Movilidad para Profesores de las Universidades Públicas de Madrid 2011–2012 to L.delP); Canadian Institutes of Health Research (CIHR) [MOP-82875 to W.W.W.]; Natural Sciences and Engineering Research Council of Canada (NSERC) [RGPIN355532-10 to W.W.W.]; National Institutes of Health [1R01GM084875 toW.W.W.]; CSIC (Spanish National Research Council) [JAE-Doc grant-2010 to O.R., in part by the European Social Fund]. Spanish science, technology and innovation contract [University of Castilla-LaMancha-2014 to O.R., in part by the European Social Fund]. Funding for open access charge: MICINN [SAF2011 24225 to L.delP., SAF2014-53819-R to L.delP., B.J.

Glucocorticoids Antagonize Ap-1 by Inhibiting the Activation/Phosphorylation of Jnk without Affecting Its Subcellular Distribution

The immunosuppressive and antiinflammatory actions of glucocorticoid hormones are mediated by their transrepression of activating protein-1 (AP-1) and nuclear factor-kappa B (NFκB) transcription factors. Inhibition of the c-Jun NH2-terminal kinase (JNK) signaling pathway, the main mediator of AP-1 activation, has been described in extracts of hormone-treated cells. Here, we show by confocal laser microscopy, enzymatic assays, and immunoblotting that the synthetic glucocorticoid dexamethasone inhibited tumor necrosis factor α (TNF-α)–induced phosphorylation and activation of JNK in the cytoplasm and nucleus of intact HeLa cells. As a result, c-Jun NH2-terminal domain phosphorylation and induction were impaired. Dexamethasone did not block the TNF-α–induced JNK nuclear translocation, but rather induced, per se, nuclear accumulation of the enzyme. Consistently with previous findings, a glucocorticoid receptor mutant (GRdim), which is deficient in dimerization, DNA binding, and transactivation, but retains AP-1 transrepressing activity, was as efficient as wild-type GR in mediating the same effects of dexamethasone on JNK in transfected Cos-7 cells. Our results show that glucocorticoids antagonize the TNF-α–induced activation of AP-1 by causing the accumulation of inactive JNK without affecting its subcellular distribution

Nuclear DICKKOPF-1 as a biomarker of chemoresistance and poor clinical outcome in colorectal cancer

Sporadic colorectal cancer (CRC) insurgence and progression depend on the activation of Wnt/β-catenin signaling. Dickkopf (DKK)-1 is an extracellular inhibitor of Wnt/β-catenin signaling that also has undefined β-catenin-independent actions. Here we report for the first time that a proportion of DKK-1 locates within the nucleus of healthy small intestine and colon mucosa, and of CRC cells at specific chromatin sites of active transcription. Moreover, we show that DKK-1 regulates several cancer-related genes including the cancer stem cell marker aldehyde dehydrogenase 1A1 (ALDH1A1) and Ral-binding protein 1-associated Eps domain-containing 2 (REPS2), which are involved in detoxification of chemotherapeutic agents. Nuclear DKK-1 expression is lost along CRC progression; however, it remains high in a subset (15%) of CRC patients (n = 699) and associates with decreased progression-free survival (PFS) after chemotherapy administration and overall survival (OS) [adjusted HR, 1.65; 95% confidence interval (CI), 1.23-2.21; P = 0.002)]. Overexpression of ALDH1A1 and REPS2 associates with nuclear DKK-1 expression in tumors and correlates with decreased OS (P = 0.001 and 0.014) and PFS. In summary, our findings demonstrate a novel location of DKK-1 within the cell nucleus and support a role of nuclear DKK-1 as a predictive biomarker of chemoresistance in colorectal cancer

Hypoxia Negatively Regulates Antimetastatic PEDF in Melanoma Cells by a Hypoxia Inducible Factor-Independent, Autophagy Dependent Mechanism

Pigment epithelium-derived factor (PEDF), a member of the serine protease inhibitor (SERPIN) superfamily, displays a potent antiangiogenic and antimetastatic activity in a broad range of tumor types. Melanocytes and low aggressive melanoma cells secrete high levels of PEDF, while its expression is lost in highly aggressive melanomas. PEDF efficiently abrogates a number of functional properties critical for the acquisition of metastatic ability by melanoma cells, such as neovascularization, proliferation, migration, invasiveness and extravasation. In this study, we identify hypoxia as a relevant negative regulator of PEDF in melanocytes and low aggressive melanoma cells. PEDF was regulated at the protein level. Importantly, although downregulation of PEDF was induced by inhibition of 2-oxoglutarate-dependent dioxygenases, it was independent of the hypoxia inducible factor (HIF), a key mediator of the adaptation to hypoxia. Decreased PEDF protein was not mediated by inhibition of translation through untranslated regions (UTRs) in melanoma cells. Degradation by metalloproteinases, implicated on PEDF degradation in retinal pigment epithelial cells, or by the proteasome, was also excluded as regulatory mechanism in melanoma cells. Instead, we found that degradation by autophagy was critical for PEDF downregulation under hypoxia in human melanoma cells. Our findings show that hypoxic conditions encountered during primary melanoma growth downregulate antiangiogenic and antimetastasic PEDF by a posttranslational mechanism involving degradation by autophagy and could therefore contribute to the acquisition of highly metastatic potential characteristic of aggressive melanoma cells

Policy Recommendations ebook. Migrations, Gender and Inclusion from an International Perspective

Technical coordination: Rais, Carlo (Pixel Associazione - Italy); Martellini, Lorenzo (Pixel Associazione - Italy) Collaborating team: Esteban-Ibañez, Macarena (Pablo de Olavide University - Spain); Macias Gómez-Esthern, Beatriz (Pablo de Olavide University - Spain) Monreal Gimeno, Carmen (Pablo de Olavide University - Spain) Moreno Amador, Gracia (Pablo de Olavide University - Spain) Pérez-de-Guzmán-Puya, Victoria (Pablo de Olavide University - Spain)This publication is the third product of the Erasmus + Project entitled Voices of Immigrant Women (Project Number: 2020-1-ES01-KA203-082364). This product is based on a set of policy recommendations that provides practical guidance on intervention proposals to those with political responsibilities in governance on migration management and policies for integration and social inclusion, as well as to policy makers in the governance of training in Higher Education (University) at all levels. This is intended to promote the development of practical strategies that allow overcoming the obstacles encountered by migrant women during the integration process, favoring the construction of institutions, administrations and, ultimately, more inclusive societies. The content presented in this book proposes recommendations and intervention proposals oriented to practice to: - Improve Higher Education study plans by promoting the training of students as future active protagonists who are aware of social interventions. This will promote equity, diversity and the integration of migrant women. - Strengthen cooperation and creation of networks between academic organizations, the third sector and public administrations that are responsible for promoting the integration and inclusion of migrant women. - Promote dialogue and the exchange of knowledge to, firstly, raise awareness of human mobility and gender in Europe and, secondly, promote the participation and social, labor and civic integration of the migrant population. All this is developed through 4 areas in which this book is articulated. The first area entitled "Migrant women needs and successful integration interventions"; the second area entitled "Promoting University students awareness and civic and social responsibility towards migrant women integration"; the third area entitled "Cooperation between Higher Education institutions and third sector"; the fourth and last area, entitled "Inclusive Higher Education".PixelInstitut De Recherche Pour Le DéveloppementPolytechnic Institute of BragançaUniversity of FlorenceEuropean Public Law OrganizationThe Peace InstituteEmet Arco Iris FoundationUniversidad Pablo de Olavid

Mecanismo de inhibición de la angiogénesis tumoral por trombospondina-1

Peer reviewe

lnc RNAs, hypoxia and metastasis

In a variety of pathophysiological conditions oxygen demand exceeds its supply, leading to a compromising condition referred to as hypoxia, which triggers complex adaptive mechanisms mediated primarily through transcriptional reprogramming by hypoxia inducible factors (HIFs). Solid tumors are an example of this condition, as deregulated proliferation generates oxygen deficient microenvironments to which cancer cells need to adapt. Among the adaptive processes, the growth of new vessels (angiogenesis) and metabolic reprogramming are central to cope with low oxygen tension. However, in the context of solid tumors, hypoxic areas perpetuate and expand due to the aberrant activation of angiogenesis which gives rise to abnormal tumor vessels and a dysfunctional tumor vasculature. In fact, the prognostic significance of hypoxic regions is well recognized and tumor hypoxia is causally associated with resistance to therapy. Additionally, hypoxia induces metastasis through regulation of several critical processes implicated in the complex metastatic cascade, especially epithelial-mesenchymal transition and cancer stem cell renewal. Recent studies have implicated HIF target genes in nearly every step of the metastatic process; however, the precise molecular and cellular mechanisms underlying hypoxia-induced metastatic dissemination are just beginning to be uncovered.Peer Reviewe