Control of genomic stability by APC/C-Cdh1 and therapeutic implications

Tesis doctoral inédita, leída en Universidad Autónoma de Madrid, facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 11-11-2013The E3-­‐ubiquitin ligase APC/C-­‐Cdh1 is essential for embryonic endoreduplication but its relevance in the mammalian mitotic cell cycle is still unclear. We show here that genetic ablation of Cdh1 in the developing nervous system results in hypoplastic brain, abnormal development of ependymal cells and hydrocephalus. These defects correlate with increased levels of multiple cell cycle regulators and increased entry into S-­‐phase in neural progenitors, resulting in replicative stress. However, cell division is prevented in the absence of Cdh1 due to the activation the DNA damage response, induction of p53, G2 arrest and apoptotic death. Concomitant ablation of p53 rescues apoptosis but not replicative stress, resulting in premature death due to the presence of damaged neurons throughout the adult brain. Interestingly, partial inhibition of cyclin-­‐dependent kinases (Cdks) rescues the replicative stress and the defective proliferation suggesting that Cdh1 loss results in DNA-­‐damage-­‐like response due to Cdk hyperactivation. In addition, by using a proteomic approach in Cdh1-­‐null cells and mouse tissues, we have identified the kinesin Eg5 and topoisomerase 2α as APC/C-­‐Cdh1 targets involved in the maintenance of genomic stability. The high levels of Eg5 in Cdh1-­‐null cells are accompanied by partial resistance to Eg5 inhibitors such as monastrol. In contrast, Cdh1-­‐null cells display a dramatic sensitivity to Top2α poisons currently used in cancer therapy as a consequence of increased levels of trapped Top2α-­‐DNA complexes. Treatment of human cancer cells with APC/C inhibitors results in increased sensitivity to Top2α poisons revealing a new synthetic lethal interaction that could be used for the optimization of anticancer treatments. These data indicate that a) Inactivation of Cdh1 in vivo results in replicative stress, cell cycle arrest and cell death; and b) APC/C inhibition may have therapeutic use, not only by inhibiting Cdc20 leading to mitotic arrest, but also by altering the levels of Cdh1 substrates (such as Eg5 and Top2α). Thus, targeting the APC/C may result in differential responses (increased resistance or susceptibility) to specific therapeutic agents.La E3-­‐ubiquitina ligasa APC/C-­‐Cdh1 es esencial para la endoreduplicación durante el desarrollo embrionario, pero su relevancia en el ciclo celular mitótico de mamíferos todavía no está clara. La eliminación genética de Cdh1 específicamente en el sistema nervioso resulta en hipoplasia, desarrollo anormal de las células ependimarias e hidrocefalia. Estos defectos correlacionan con un incremento en niveles de reguladores del ciclo celular y en entrada en fase S en los progenitores neurales, que da lugar a estrés replicativo. En ausencia de Cdh1 estas células progenitoras no progresan en el ciclo debido a la activación de la respuesta a daño en el DNA, la inducción de p53, la parada en la fase G2 y la muerte celular programada (apoptosis). La eliminación de p53 previene la apoptosis de las células deficientes para Cdh1, pero no el estrés replicativo. Sin embargo, la inhibición parcial de la actividad de quinasas dependientes de ciclinas (Cdks) rescata el estrés replicativo y los problemas en proliferación, lo que sugiere que la acumulación de daño en el DNA en ausencia de Cdh1 se debe a la hiperactivación de las Cdks. Además, mediante técnicas proteómicas en células y tejidos deficientes para Cdh1, hemos identificado a la kinesina Eg5 y la topoisomerasa 2α como nuevos sustratos de APC/C-­‐Cdh1 implicados en el mantenimiento de la estabilidad genómica. La acumulación de Eg5 en células donde se ha eliminado Cdh1 provoca una resistencia parcial al inhibidor de Eg5 monastrol. Por otro lado, las células deficientes para Cdh1 son especialmente sensibles al inhibidor de topoisomerasa etopósido, actualmente utilizado en tratamientos antitumorales. De hecho, el tratamiento de líneas tumorales humanas con un inhibidor de APC/C incrementa la sensibilidad a inhibidores de topoisomerasa 2. Estos datos muestran que: a) La inactivación de Cdh1 in vivo provoca estrés replicativo, parada del ciclo y muerte celular; and b) La inhibición de APC/C puede tener aplicaciones terapéuticas, no sólo a través de la inducción de parada en mitosis por bloqueo de la actividad de cdc20, sino también mediante el aumento de niveles de sustratos de Cdh1 (tales como Eg5 o Top2α) que modulan la respuesta a distintos tratamientos contra el cancer

Valuation of differences between sports specialization and multisport practice using FMS in soccer players : a transversal study

RESUMEN: La batería de pruebas llamada functional movement screen (FMS) permite valorar la calidad de patrones de movimiento fundamentales para identificar limitaciones individuales o asimetrías. Objetivo: El objetivo de este estudio fue comparar la puntuación en la batería de pruebas FMS entre deportistas especializados en una modalidad deportiva y aquellos que han practicado más de una modalidad deportiva en el pasado. Material y métodos: En el estudio han participado un total de 13 jugadores de fútbol (5 especializados y 8 multideporte) pertenecientes a la tercera división cántabra con edades comprendidas entre los 18 y 32 años. Se llevaron a cabo las siete pruebas funcionales recibiendo en cada una de ellas una puntuación de entre 0 y 3 puntos haciendo referencia a los parámetros observados por el examinador. Resultados: Los resultados obtenidos muestran que entre grupos observamos diferencias que podemos clasificar como triviales en el caso del in line lunge (d = -0.303), active straight leg raise (d = -0.047) y trunk stability push up (d = 0.080), en concreto estas diferencias son a favor del grupo especializado en trunk stability push up, y a favor del grupo multideporte en las otras dos. También existen diferencias pequeñas en deep squat (d = 0.229), a favor del grupo especializado así como en shoulder mobilty (d = 0.496) y rotatory stability (d = 0.396) y moderadas en el hurdle step (d = 0.720), a favor del grupo especializado de nuevo. Discusión: La conclusión final hace referencia a que no existen diferencias significativas entre ambos grupos en cuanto a la puntuación total de la batería de pruebas. Sin embargo, si observamos la magnitud de estas diferencias encontramos diferentes resultados dependiendo de qué prueba estemos valorando.ABSTRACT: The battery of functional tests called functional movement screen (FMS) allows to assess the quality of fundamental movement patterns to identify individual limitations or asymmetries. Objective: The objective of this study was to compare the score in the FMS battery tests between athletes specialized in a sport modality and those who have practiced more than one sport modality in the past. Material and methods: A total of 13 soccer players (5 specialized and 8 multisport) belonging to the Cantabrian third division with ages between 18 and 32 years participated in the study. The seven functional tests were carried out receiving a score between 0 and 3 each one, referring to the parameters observed by the examiner. Results: The results obtained show that between groups we observe differences classified as trivial in the case of in line lunge, active straight leg raise and trunk stability push up (d <0.2), specifically these differences are in favor of the specialized group in trunk stability push up, and in favor of multisport group in the other two. There is also small differences in deep squat, in favor of the specialized group as well as shoulder mobilty and rotatory stability (d = 0.2-0.59) and moderate differences in the hurdle step (d = 1.20-1.99), in favor of the specialized group again. Discussion: The final conclusion refers to the fact that there are no significant differences between both groups regarding the total score of the battery of tests. However, the results obtained in certain tests in relation to the magnitude of these differences are interesting, as we can see in the hurdle step.Grado en Fisioterapi

The APC/C cofactor Cdh1 prevents replicative stress and p53-dependent cell death in neural progenitors

The E3-ubiquitin ligase APC/C-Cdh1 is essential for endoreduplication but its relevance in the mammalian mitotic cell cycle is still unclear. Here we show that genetic ablation of Cdh1 in the developing nervous system results in hypoplastic brain and hydrocephalus. These defects correlate with enhanced levels of Cdh1 substrates and increased entry into the S phase in neural progenitors. However, cell division is prevented in the absence of Cdh1 due to hyperactivation of cyclin-dependent kinases, replicative stress, induction of p53, G2 arrest and apoptotic death of these progenitor cells. Concomitant ablation of p53 rescues apoptosis but not replicative stress, resulting in the presence of damaged neurons throughout the adult brain. These data indicate that the inactivation of Cdh1 in vivo results in replicative stress, cell cycle arrest and cell death, supporting recent therapeutic proposals aimed to inhibit the APC/C in tumours.M.E. was supported by the Spanish Ministry of Economy and Competitiveness (MINECO). This work was funded by grants from the Foundation Ramón Areces and MINECO SAF2012-38215 to M.M.). The Cell Division and Cancer Group of the CNIO are supported by the OncoCycle Programme (S2010/BMD-2470) from the Comunidad de Madrid, the OncoBIO Consolider-Ingenio 2010 Programme (MINECO, CSD2007-00017) and the European Union Seventh Framework Programme (MitoSys project; HEALTH-F5-2010-241548).Peer Reviewe

Maritime studies and shipping bussines: a trend research on education programs

The Schools of Maritime Engineering & Business of the Spanish Universities forms the future professionals of the Spanish Merchant Marine who will sail the seven seas transporting goods in all its various forms. This remains a very important profession however, the importance of these activities it is not usually well known by the rest of the society not directly involved with the maritime world. The professions have been evolving, and the maritime job is not an exception. The marine professionals had opened new gates to develop new activities in other di erent positions which traditionally were busy by others carriers, for example, shipping companies. However, training in this sense is lacking, nautical students in most of the cases are not trained and specialized in this regard. The idea of this research paper is to quantify and identify the needs for the training and propose a training plan according to current needs in this area. The methodology of this work, has consisted of comparative analysis based on the results obtained from di erent surveys. The first survey has been conducted in several schools of nautical and maritime studies, considering the di erent levels existent at training programs, perception from students in relation to major weaknesses and missing elements. The second survey focused on experts in shipping & maritime logistics considering their experience from the trainee students with the objective to identify the core skills that will be required from a professional point of view. The results of both surveys have been combined, performing a comparative analysis in order to define a curriculum model that should cover the academic and business abilities gaps identified during the study

Targeting Mitotic Exit Leads to Tumor Regression In Vivo: Modulation by Cdk1, Mastl, and the PP2A/B55α,δ Phosphatase

SummaryTargeting mitotic exit has been recently proposed as a relevant therapeutic approach against cancer. By using genetically engineered mice, we show that the APC/C cofactor Cdc20 is essential for anaphase onset in vivo in embryonic or adult cells, including progenitor/stem cells. Ablation of Cdc20 results in efficient regression of aggressive tumors, whereas current mitotic drugs display limited effects. Yet, Cdc20 null cells can exit from mitosis upon inactivation of Cdk1 and the kinase Mastl (Greatwall). This mitotic exit depends on the activity of PP2A phosphatase complexes containing B55α or B55δ regulatory subunits. These data illustrate the relevance of critical players of mitotic exit in mammals and their implications in the balance between cell death and mitotic exit in tumor cells

A Social-Emotional Learning Program for Suicide Prevention through Animal-Assisted Intervention

The aim of the study was to carry out a pilot implementation and evaluation of the OverCome-AAI program, a pioneering program for the prevention of suicidal behavior through animal-assisted interventions for young people with high risk factors for suicidal behavior. The study sample consisted of 30 adolescents (11 boys and 19 girls) aged between 14 and 17 years (Mean age = 15.50, SD = 1.60) from the Basque Country (Northern Spain). After the intervention, subjects presented reductions in suicidal ideation, suicide plans, and non-suicidal self-harm, as well as a greater predisposition to seek help. A reduction in the intensity of mental pain was also found, although no differences were observed in indicators of hopelessness and depression. The preliminary results obtained in this pilot study suggest that the OverCome-AAI program may be effective in reducing suicidal behavior and non-suicidal self-harm in young people in residential care who present high risk factors for suicide.This research was funded by the call for research grants at the UPV/EHU 2017, grant number (US18/05), Agintzari Sociedad Cooperativa de Iniciativa Social, and by the call for research grants at the Basque Government, grant number (IT892-16)

Cloud Robotics: Sistema Multi-Robot conectado al Cloud público AWS

Este trabajo presenta el despliegue de un escenario de sistema multi-robot, integrado por robots conectados vía WiFi al Cloud público de Amazon Web Services, simulando un sistema de riego artificial de campo. Se detallan los prototipos desarrollados, como así también el protocolo de comunicación utilizado con el servicio de AWS IoT, y los algoritmos implementados que permiten procesar las imágenes adquiridas e identificar determinados patrones y objetos.XVI Workshop Procesamiento Distribuido y Paralelo (WPDP).Red de Universidades con Carreras en Informática (RedUNCI

Cloud Robotics: Sistema Multi-Robot conectado al Cloud público AWS

Este trabajo presenta el despliegue de un escenario de sistema multi-robot, integrado por robots conectados vía WiFi al Cloud público de Amazon Web Services, simulando un sistema de riego artificial de campo. Se detallan los prototipos desarrollados, como así también el protocolo de comunicación utilizado con el servicio de AWS IoT, y los algoritmos implementados que permiten procesar las imágenes adquiridas e identificar determinados patrones y objetos.XVI Workshop Procesamiento Distribuido y Paralelo (WPDP).Red de Universidades con Carreras en Informática (RedUNCI

Intelligent Distributed System for Energy Efficient Control

In this work, we present an intelligent system developed for energy consumption distributed control and monitoring. It supports real time cloudbased data visualization of power profiles from different areas, so as to optimize overall power consumption. The local intelligent processing unit (LIPU) that control the different environments is described. The communication network model that allows connecting multiple LIPUs to apply power consumption policies defined by the organization is analyzed, and the unit's capabilities in relation to cloud connectivity and realtime processing are considered through a theoretical scalability study. Finally, we describe relevant implementation features in the context of “Facultad de Informática” of the “Universidad Nacional de La Plata” (Argentine).Instituto de Investigación en Informátic

Cloud Robotics: Sistema Multi-Robot conectado al Cloud público AWS

Este trabajo presenta el despliegue de un escenario de sistema multi-robot, integrado por robots conectados vía WiFi al Cloud público de Amazon Web Services, simulando un sistema de riego artificial de campo. Se detallan los prototipos desarrollados, como así también el protocolo de comunicación utilizado con el servicio de AWS IoT, y los algoritmos implementados que permiten procesar las imágenes adquiridas e identificar determinados patrones y objetos.XVI Workshop Procesamiento Distribuido y Paralelo (WPDP).Red de Universidades con Carreras en Informática (RedUNCI