Estudio de la metilación de los genes FLNc y POLE en cáncer de mama

[ES] La epigenética comprende el estudio de modificaciones en la expresión de genes que no obedecen a alteraciones en la secuencia de ADN. Entre estas modificaciones se encuentra la metilación del ADN, que puede alterar anormalmente la expresión de genes en procesos celulares y contribuir al desarrollo y progresión del cáncer de mama. Las células tumorales se caracterizan por una pérdida masiva de metilación y al mismo tiempo por la adquisición de un patrón de hipermetilación en islas CpG de ciertos promotores. En el caso de genes reparadores del ADN, ciertas metilaciones anormales podrían causar un silenciamiento del gen, por lo que se dejaría de realizar correctamente esta función reparadora. Alteraciones en los niveles normales de metilación de estos genes podrían estar implicadas en la etiología del tumor, así como en los mecanismos de resistencia a alguno de los tratamientos actuales. Por otra parte, los genes codificantes de proteínas de adhesión y estructura celular desempeñan un papel fundamental en la progresión y metástasis, debido a que pueden facilitar la interacción de las células tumorales con células de tejidos lejanos. De la misma manera, ciertas modificaciones en estos genes podrían relacionarse con diferencias en la capacidad de los tumores de producir una metástasis. En este proyecto se ha estudiado la metilación de regiones concretas de POLE, un gen reparador del ADN; y FLNc, gen relacionado con la estructura y adhesión celular, mediante la tecnología Sequenom. Se han utilizado líneas celulares de diferentes subtipos de cáncer de mama (HER2+, luminales y triple negativo) y las líneas no tumorales de mama MCF10A y MCF12A, con el objetivo de encontrar un posible patrón de metilación relacionado con la existencia de la enfermedad y su desarrollo y por lo tanto, un posible marcador de diagnóstico y/o pronóstico. Esto es de especial importancia en el subtipo triple negativo, el cual actualmente carece de un tratamiento anti- diana específico efectivo.[EN] Epigenetics comprise the study of gene expression modifications different from DNA sequence alterations. One of this modifications is DNA methylation, which may alter gene expression in cellular processes and contribute to breast cancer development and progression. Tumour cells are characterized by a massive loss of methylation and also a hypermethylated pattern in certain promoters. In DNA repair genes, certain abnormal methylations may cause gene silencing, and therefore damaged DNA would not be repaired. Alterations in methylation levels may be involved in tumour etiology and also in some resistance mechanisms of current treatments. Structural and cell adhesion proteins coding genes have a pivotal role in metastasis and progression, since it is facilitated by the interaction between tumor cells and distant tissue cells. In the same way as before, certain modifications in these genes may be related with differences in levels of metastization. In this project, methylation of certain regions of a DNA repair gene (FLNc) and a gene involved in the cell structure and adhesion (POLE) have been studied by means of Sequenom technology. Cell lines from different breast cancer subtypes (HER2+, luminal and triple negative) and the non-tumoral cell lines MCF10A and MCF12A have been used in order to find a methylation pattern related with the presence and development of breast cancer, and therefore a possible diagnostic or prognosis marker. This is particularly important in triple negative cancer, which lacks of an effective specific anti-target treatment.García Serra, R. (2017). Estudio de la metilación de los genes FLNc y POLE en cáncer de mama. http://hdl.handle.net/10251/86554TFG

Molecular characterization and clinical impact of TMPRSS2-ERG rearrangement on prostate cancer: comparison between FISH and RT-PCR

Prostate cancer (PCa) is a very heterogeneous disease, and there are constraints in its current diagnosis. Serum PSA levels, digital rectal examination (DRE), and histopathologic analysis often drive to overdiagnosis and overtreatment. Since 2005, the presence of the genetic rearrangement between transmembrane-serine protease gene (TMPRSS2) and the erythroblast transformation-specific (ETS)member ERG (v-ets erythroblastosis virus E26 oncogene homolog avian) has been demonstrated in almost half of PCa cases. Both FISH and RT-PCR are useful tools for detecting these rearrangements, but very few comparatives between both techniques have been published. In this study, we included FFPE tumors from 294 PCa patients treated with radical prostatectomy with more than 5 years of followup.We constructed a total of 20 tissue microarrays in order to perform break-apart and tricolor probe FISH approaches that were compared with RT-PCR, showing a concordance of 80.6% ( P < 0.001). The presence of TMPRSS2-ERG rearrangement was observed in 56.6% of cases. No association between TMPRSS2-ERG status and clinicopathological parameters nor biochemical progression and clinical progression free survival was found. In conclusion, this study demonstrates that both FISH and RT-PCR are useful tools in the assessment of the TMPRSS2-ERG fusion gene status in PCa patients and that this genetic feature per se lacks prognostic value.This study has been funded by the Grants FIS PI06/01619 and PI10/01206 from the Instituto de Salud Carlos III, Madrid, ACOMP 12/029 from the Generalitat Valenciana, Valencia, and Astra Zeneca, Spain

Observations in the Spanish Mediterranean Waters: A Review and Update of Results of 30-Year Monitoring

The Instituto Espa&ntilde;ol de Oceanograf&iacute;a (IEO, Spanish Institute of Oceanography) has maintained different monitoring programs in the Spanish Mediterranean waters (Western Mediterranean) since 1992. All these monitoring programs were unified in 2007 under the current program RADMED (series temporales de datos oceanogr&aacute;ficos en el Mediterr&aacute;neo), which is devoted to the in situ multidisciplinary sampling of the water column of coastal and open-sea waters by means of periodic oceanographic campaigns. These campaigns, together with a network of tide-gauges, are part of the IEO Observing system (IEOOS). In some cases, the temperature and salinity time series collected in the frame of these monitoring programs are now more than 30 years long, whereas sea level time series date to the beginning of the 1940s. This information has been complemented with international databases and has been analyzed in numerous works by the Grupo mediterr&aacute;neo de Cambio Clim&aacute;tico (GCC; Mediterranean Climate Change Group) for more than 20 years. These works have been devoted to the detection and quantification of the changes that climate change is producing on the physical, chemical, and biological properties of the Spanish Mediterranean waters. In this work, we review the results obtained by the GCC since 2005 in relation to the changes in the physical properties of the sea: water column temperature, salinity, and density, heat content, mixed layer depth, and sea level. Time series and results are updated from the last works, and the reliability of the existing time series for the detection of climatologies and long-term trends are analyzed. Furthermore, the different sources of uncertainty in the estimation of linear trends are considered in the present work. Besides this review and update of the results obtained from the data collected in the frame of the IEOOS, we conduct a review of the existing monitoring capabilities from other institutions in the Spanish Mediterranean waters and a review of results dealing with climate change in the Spanish Mediterranean obtained by such institutions. In particular, we include a review of the results obtained by SOCIB (Servicio de Observaci&oacute;n y Predicci&oacute;n Costero de las Islas Baleares; Balearic Islands costal observing and forecasting system) in relation to the study of marine heat waves and the warming of the sea surface, and the results corresponding to the intense warming of the Catalan continental shelf at L&rsquo;Estartit oceanographic station. All these results evidence that the surface Spanish Mediterranean waters are warming up at a rate higher than that affecting the global ocean (&gt;2 &deg;C/100 years). This warming and a salinity increase are also observed along the whole water column. Marine heat waves are increasing their intensity, frequency, and duration since 1982, and coastal sea level is increasing at a rate of 2.5 mm/yr. The salinity increase seems to have compensated for the warming, at least at surface and intermediate waters where no significant trends have been detected for the density. This could also be the reason for the lack of significant trends in the evolution of the mixed layer depth. All these results highlight the importance of monitoring the water column and the necessity of maintaining in situ sampling programs, which are essential for the study of changes that are occurring throughout the Spanish Mediterranean waters

Malignant Arrhythmogenic Role Associated with RBM20: A Comprehensive Interpretation Focused on a Personalized Approach

The RBM20 gene encodes the muscle-specific splicing factor RNA-binding motif 20, a regulator of heart-specific alternative splicing. Nearly 40 potentially deleterious variants in RBM20 have been reported in the last ten years, being found to be associated with highly arrhythmogenic events in familial dilated cardiomyopathy. Frequently, malignant arrhythmias can be a primary manifestation of disease. The early recognition of arrhythmic genotypes is crucial in avoiding lethal episodes, as it may have an impact on the adoption of personalized preventive measures. Our study performs a comprehensive update of data concerning rare variants in RBM20 that are associated with malignant arrhythmogenic phenotypes with a focus on personalized medicine.This work was supported by Obra Social "La Caixa Foundation" (LCF/PR/GN16/50290001 and LCF/PR/GN19/50320002), Fondo Investigacion Sanitaria (FIS PI16/01203 and FIS, PI17/01690) from Instituto Salud Carlos III (ISCIII), and "Fundacio Privada Daniel Bravo Andreu". CIBERCV is an initiative of the ISCIII, Spanish Ministry of Economy and Competitiveness

The RADMED monitoring program as a tool for MSFD implementation: toward an ecosystem based approach

In the western Mediterranean Sea, the RADMED monitoring programme is already conducting several of the evaluations required under the Marine Strategy Framework Directive (MFSD) along the Spanish Mediterranean coast. The different aspects of the ecosystem that are regularly sampled under this monitoring programme are the physical environment and the chemical and biological variables of the water column, together with the planktonic communities, biomass and structure. Moreover, determinations of some anthropogenic stressors on the marine environment, such as contaminants and microplastics, are under development. Data are managed and stored at the Instituto Español de Oceanografía (IEO) Data Centre that works under the Sea- DataNet infrastructure, and are also stored in the IBAMar database. In combination with remote sensing data, they are used to address open questions on the ecosystems in the western Mediterranean Sea.Postprint2,293

Proyecto Escritiva

Memoria ID-0132. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2015-2016

Rare Variants Associated with Arrhythmogenic Cardiomyopathy: Reclassification Five Years Later.

Genetic interpretation of rare variants associated with arrhythmogenic cardiomyopathy (ACM) is essential due to their diagnostic implications. New data may relabel previous variant classifications, but how often reanalysis is necessary remains undefined. Five years ago, 39 rare ACM-related variants were identified in patients with features of cardiomyopathy. These variants were classified following the American College of Medical Genetics and Genomics' guidelines. In the present study, we reevaluated these rare variants including novel available data. All cases carried one rare variant classified as being of ambiguous significance (82.05%) or likely pathogenic (17.95%) in 2016. In our comprehensive reanalysis, the classification of 30.77% of these variants changed, mainly due to updated global frequencies. As in 2016, nowadays most variants were classified as having an uncertain role (64.1%), but the proportion of variants with an uncertain role was significantly decreased (17.95%). The percentage of rare variants classified as potentially deleterious increased from 17.95% to 23.07%. Moreover, 83.33% of reclassified variants gained certainty. We propose that periodic genetic reanalysis of all rare variants associated with arrhythmogenic cardiomyopathy should be undertaken at least once every five years. Defining the roles of rare variants may help clinicians obtain a definite diagnosis

Diseño y desarrollo de un modelo de portafolio para la evaluación de competencias en el Grado de Estudios Portugueses y Brasileños

Memoria ID-0205. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015