Estudio de los perfiles de expresión de microRNAs en líneas celulares de cáncer de mama triple negativo tratadas con doxorubicina: implicación de la familia miR-449.

Entre los diferentes subtipos de cáncer de mama se encuentra el triple negativo, denominado así porque las células tumorales de este subtipo presentan baja expresión o carecen de receptores de estrógenos, receptores de progesterona, y receptores del factor de crecimiento epidérmico tipo II (HER2). Actualmente no existe una diana específica para el tratamiento de este tipo de cáncer de mama, caracterizado por ser muy agresivo, presentar alto riesgo de metástasis, un alto índice de recurrencia, y una bajo índice de supervivencia. Debido a la falta de dianas moleculares específicas, el tratamiento comúnmente utilizado es la quimioterapia. Dentro de los diferentes agentes quimioterápicos, se encuentra la doxorubicina, una antraciclina que produce disrupción del DNA y muerte celular. Sin embrago, un gran porcentaje de las pacientes triple negativas tratadas con antraciclinas presentan enfermedad residual en los tres años posteriores al tratamiento, además de un incremento en el riesgo de mortalidad. Los mecanismos de quimiosensibilidad y quimioresistencia a la doxorubicina todavía no están claros, por lo que el estudio de las vías de regulación y posibles dianas específicas podría optimizar la respuesta a este tratamiento. En este sentido, los microRNAs (miRNAs) surgen como un nuevo peldaño de regulación del cáncer y respuesta a tratamiento. En este trabajo de investigación analizamos los niveles de expresión de miRNAs en líneas celulares de cáncer de mama con subtipo molecular triple negativo (MDA-MB-231 y MDA-MB-468) y luminal A (MCF-7), tras el tratamiento con doxorubicina. Los resultados nos permiten establecer grupos de miRNAs que cambian tras el tratamiento y que pueden predecir una respuesta diferencial al fármaco. Además, mediante herramientas bioinformáticas, estudiamos la posible interacción entre los miRNAs y sus potenciales dianas. Finalmente nos centramos en la línea celular triple negativa MDA-MB-231 y en la familia de miRNAs-449, puesto que todos sus miembros se sobre-expresan tras el tratamiento. De manera paralela trabajamos con una línea celular triple negativa resistente a doxorubicina (MDA-MB-231R) donde medimos la expresión tanto de la familia de miRNAs-449 como de sus dianas, y encontramos diferencias significativas respecto de las líneas triples negativas sensibles al fármaco. Las pruebas funcionales (tanto de sobreexpresión como de inhibición de esos microRNAs) demuestran que esta familia regula vías de respuesta al fármaco relacionadas con apoptosis y/o ciclo celular a través de la interacción con sus dianas génicas. Por otra parte, realizamos estudios de viabilidad y ciclo celular, con el objetivo de analizar el posible papel de estos miRNAs en la respuesta a la doxorubicina a través de la des-regulación del ciclo celular. Además medimos la expresión de esta familia de miRNAs y de sus dianas en pacientes de cáncer de mama triple negativo tratadas con antraciclinas. En conjunto, los resultados obtenidos indican que esta familia de miRNAs puede mediar, al menos en parte, la respuesta al fármaco, y que su sobre- o infra- expresión conducen a cambios en la sensibilidad o la resistencia al tratamiento respectivamente

Desarrollo de salsas con microalgas

[EN] Sauces are one of the food consume products that have undergone further development in the last years regarding the type of formats and flavours available. This is a very competitive market that is always looking for new flavours and colours that make them attractive. In the other hand there is a preference to use natural ingredients that provide high value-added features to the product. Microalgae are one of those ingredients because of their content in polyunsaturated fatty acids, protein profile and colour. In this thesis "Development sauces microalgae" he has worked on the development of mayonnaise-type sauces with microalgae. In the first stage we proceeded to study physicochemical parameters of a commercial mayonnaise, in order to make mayonnaise as similar to that existing in the market. In the next steep we proceeded to the formulation, preparation and subsequent assessment of physicochemical and sensory characteristics of a mayonnaise with two varieties of microalgae Spirulina maxima and Tetraselmis chuii. It is made two formulations: one mayonnaise with Spirulina maxima and the other one with Tetraselmis chuii. Getting two mayonnaises with natural colours from microalgae, it depends on its rate in the formulation gives them different intensity of colour making green the mayonnaise with Tetraselmis and blue the mayonnaise with Spirulina, besides increasing the mayonnaise nutritional content and also giving a characteristic smell and taste of sea bigger in the Tetraselmis Chuii. Finally aspects related with marketing such as product prices were addressed.[ES] El proyecto a desarrollar tiene por objetivo el desarrollo de nuevas salsas tipo mayonesa a partir de la utilización de microalgas (tetraselmis y espirulina) en su formulación. Se evaluarán las propiedades reológicas, funcionales y sensoriales de las mismas con el objetivo de la obtención de un producto que pueda ser comercializado.Tormo Llopis, JE. (2015). Desarrollo de salsas con microalgas. http://hdl.handle.net/10251/54287.TFG

An enhanced switching policy for buck-derived multi-level switching power amplifiers

This work proposes a switching policy for multi-level fullbridge switching power converters and analyses their performance by driving them with a multi-level PWM modulation, targeting highefficiency power amplifiers. Unlike conventional policies, which generate the output voltage levels only from the values of the supply voltages, this enhanced policy also uses the values of the voltage difference between supply voltages to generate additional output voltage levels, therefore maximising the number of output voltage levels for a given set of supply voltages and connection switches. Simulation results show that, when tracking a band-limited signal, the proposed switching policy can reduce the power of the high-frequency spectral content from 21% to 11% by upgrading a 5-level amplifier to a 7-level amplifier without adding supply voltages or connection switches.Peer ReviewedPostprint (published version

Targeting HER2-AXL heterodimerization to overcome resistance to HER2 blockade in breast cancer

Breast cancer; HeterodimerizationCáncer de mama; HeterodimerizaciónCàncer de mama; HeterodimeritzacióAnti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, different mechanisms play a role in treatment resistance. Here, we identified AXL overexpression as an essential mechanism of trastuzumab resistance. AXL orchestrates epithelial-to-mesenchymal transition and heterodimerizes with HER2, leading to activation of PI3K/AKT and MAPK pathways in a ligand-independent manner. Genetic depletion and pharmacological inhibition of AXL restored trastuzumab response in vitro and in vivo. AXL inhibitor plus trastuzumab achieved complete regression in trastuzumab-resistant patient-derived xenograft models. Moreover, AXL expression in HER2-positive primary tumors was able to predict prognosis. Data from the PAMELA trial showed a change in AXL expression during neoadjuvant dual HER2 blockade, supporting its role in resistance. Therefore, our study highlights the importance of targeting AXL in combination with anti-HER2 drugs across HER2-amplified breast cancer patients with high AXL expression. Furthermore, it unveils the potential value of AXL as a druggable prognostic biomarker in HER2-positive breast cancer.A.A.-A., E.J.A., and F.B.-M. were supported by Asociación Española contra el Cáncer AECC (PRDVA18013LLUC to A.A.-A., POSTD211413AREN to E.J.A., and AECC_Postdoctoral17-1062 to F.B.-M.). A.M.-S. was funded by the Spanish Government (PFIS FI20/00188). J.Ar. is supported by Breast Cancer Research Foundation (BCRF-20-08), Instituto de Salud Carlos III Project reference number AC15/00062, and the EC under the framework of the ERA-NET TRANSCAN-2 initiative cofinanced by FEDER, Instituto de Salud Carlos III (CB16/12/00449 and PI19/01181), and Asociación Española Contra el Cáncer (AECC). A.P. was supported by Instituto de Salud Carlos III—PI19/01846, Breast Cancer Now—2018NOVPCC1294. P.E. and A.L. were funded by Instituto de Salud Carlos III and cofinanced by FEDER (PI18/01219 to P.E. and CB16/12/00481 to A.L.). J.M.C. was funded by Sociedad Española de Oncología Médica (Rio Hortega-SEOM) and Compromiso ADAMED

Pruning waste management and climate change in Sierra Mágina's olive groves (Andalusia, Spain)

Unidad de excelencia María de Maeztu MdM-2015-0552In the context of climate change, concern is raising about the negative effects of some pruning waste management practices. On the one hand, burning of pruning residue is seen as controversial regarding its contribution to greenhouse gas emissions. On the other hand, chipping the wood and using it as mulch is seen as highly risky regarding pest and disease control. Considering these issues, it is important to try to understand how and why these practices are adopted. In this study, we conducted in-depth interviews and survey questionnaires in the olive-growing region of Sierra Mágina (Andalusia) in order to explore (1) which were the pruning waste management practices in place, (2) which had been these practices in the past, and (3) which were the factors influencing the choice of different practices. Since 2010, there has been a shift in pruning waste management practices in this region, from stubble burning to chipping. This change seems to be led by older/retired farmers that have young and non-inherited groves. Moreover, this change is shown not to be necessarily a result of "greening" in agriculture but rather a result of intensification and mechanization. These results are discussed regarding the processes of innovation adoption in the region and the possible unforeseen externalities that derive from this change in pruning waste management practices

Pruning waste management and climate change in Sierra Mágina's olive groves (Andalusia, Spain)

Unidad de excelencia María de Maeztu MdM-2015-0552In the context of climate change, concern is raising about the negative effects of some pruning waste management practices. On the one hand, burning of pruning residue is seen as controversial regarding its contribution to greenhouse gas emissions. On the other hand, chipping the wood and using it as mulch is seen as highly risky regarding pest and disease control. Considering these issues, it is important to try to understand how and why these practices are adopted. In this study, we conducted in-depth interviews and survey questionnaires in the olive-growing region of Sierra Mágina (Andalusia) in order to explore (1) which were the pruning waste management practices in place, (2) which had been these practices in the past, and (3) which were the factors influencing the choice of different practices. Since 2010, there has been a shift in pruning waste management practices in this region, from stubble burning to chipping. This change seems to be led by older/retired farmers that have young and non-inherited groves. Moreover, this change is shown not to be necessarily a result of "greening" in agriculture but rather a result of intensification and mechanization. These results are discussed regarding the processes of innovation adoption in the region and the possible unforeseen externalities that derive from this change in pruning waste management practices

MicroRNA profile in very young women with breast cancer

BACKGROUND: Breast cancer is rarely diagnosed in very young women (35 years old or younger), and it often presents with distinct clinical-pathological features related to a more aggressive phenotype and worse prognosis when diagnosed at this early age. A pending question is whether breast cancer in very young women arises from the deregulation of different underlying mechanisms, something that will make this disease an entity differentiated from breast cancer diagnosed in older patients. METHODS: We performed a comprehensive study of miRNA expression using miRNA Affymetrix2.0 array on paraffin-embedded tumour tissue of 42 breast cancer patients 35 years old or younger, 17 patients between 45 and 65 years old and 29 older than 65 years. Data were statistically analyzed by t-test and a hierarchical clustering via average linkage method was conducted. Results were validated by qRT-PCR. Putative targeted pathways were obtained using DIANA miRPath online software. RESULTS: The results show a differential and unique miRNA expression profile of 121 miRNAs (p-value <0.05), 96 of those with a FDR-value <0.05. Hierarchical clustering grouped the samples according to their age, but not by subtype nor by tumour characteristics. We were able to validate by qRT-PCR differences in the expression of 6 miRNAs: miR-1228*, miR-3196, miR-1275, miR-92b, miR-139 and miR-1207. Moreover, all of the miRNAs maintained the expression trend. The validated miRNAs pointed out pathways related to cell motility, invasion and proliferation. CONCLUSIONS: The study suggests that breast cancer in very young women appears as a distinct molecular signature. To our knowledge, this is the first time that a validated microRNA profile, distinctive to breast cancer in very young women, has been presented. The miRNA signature may be relevant to open an important field of research in order to elucidate the underlying mechanism in this particular disease, which in a more clinical setting, could potentially help to identify therapeutic targets in this particular set of patients.MPC is funded by the Generalitat Valenciana VALi + d, ACIF/2011/270. MTM is funded by”Rio Hortega Project” (CM12/00264). GR is a FIS “Miquel Servet” Researcher. AB holds a Translational Research Grant awarded by the Spanish Society of Medical Oncology (SEOM). This project was carried out thanks to Fundación LeCadó – proyecto Flor de Vida and co-funded by FIS project PI13/00606 and FEDER. We would like to give thanks to all the patients and volunteers for their participation and also to the INCLIVA Biobank, integrated into the Spanish Hospital Biobanks Network (ReTBioH) and supported by the Instituto de Salud Carlos III/FEDER (grant number: RD09/0076/00132). We also wish to thank several private Breast cancer associations that funded this study and the Unit for Multigenic Analysis from the Central Unit for Medical Research (UCIM/INCLIVA) for the performance of the Affymetrix microRNA profiles.S

MicroRNA-33b Suppresses Epithelial-Mesenchymal Transition Repressing the MYC-EZH2 Pathway in HER2+ Breast Carcinoma

Downregulation of miR-33b has been documented in many types of cancers and is being involved in proliferation, migration, and epithelial-mesenchymal transition (EMT). Furthermore, the enhancer of zeste homolog 2-gene (EZH2) is a master regulator of controlling the stem cell differentiation and the cell proliferation processes. We aim to evaluate the implication of miR-33b in the EMT pathway in HER2+ breast cancer (BC) and to analyze the role of EZH2 in this process as well as the interaction between them. miR-33b is downregulated in HER2+ BC cells vs healthy controls, where EZH2 has an opposite expression in vitro and in patients' samples. The upregulation of miR-33b suppressed proliferation, induced apoptosis, reduced invasion, migration and regulated EMT by an increase of E-cadherin and a decrease of ß-catenin and vimentin. The silencing of EZH2 mimicked the impact of miR-33b overexpression. Furthermore, the inhibition of miR-33b induces cell proliferation, invasion, migration, EMT, and EZH2 expression in non-tumorigenic cells. Importantly, the Kaplan-Meier analysis showed a significant association between high miR-33b expression and better overall survival. These results suggest miR-33b as a suppressive miRNA that could inhibit tumor metastasis and invasion in HER2+ BC partly by impeding EMT through the repression of the MYC-EZH2 loop

Interaction between cardiovascular risk factors and body mass index and 10-year incidence of cardiovascular disease, cancer death, and overall mortality

The effect of above-normal body mass index (BMI) on health outcomes is controversial because it is difficult to distinguish from the effect due to BMI-associated cardiovascular risk factors. The objective was to analyze the impact on 10-year incidence of cardiovascular disease, cancer deaths and overall mortality of the interaction between cardiovascular risk factors and BMI. We conducted a pooled analysis of individual data from 12 Spanish population cohorts with 10-year follow-up. Participants had no previous history of cardiovascular diseases and were 35-79years old at basal examination. Body mass index was measured at baseline being the outcome measures ten-year cardiovascular disease, cancer and overall mortality. Multivariable analyses were adjusted for potential confounders, considering the significant interactions with cardiovascular risk factors. We included 54,446 individuals (46.5% with overweight and 27.8% with obesity). After considering the significant interactions, the 10-year risk of cardiovascular disease was significantly increased in women with overweight and obesity [Hazard Ratio=2.34 (95% confidence interval: 1.19-4.61) and 5.65 (1.54-20.73), respectively]. Overweight and obesity significantly increased the risk of cancer death in women [3.98 (1.53-10.37) and 11.61 (1.93-69.72)]. Finally, obese men had an increased risk of cancer death and overall mortality [1.62 (1.03-2.54) and 1.34 (1.01-1.76), respectively]. In conclusion, overweight and obesity significantly increased the risk of cancer death and of fatal and non-fatal cardiovascular disease in women; whereas obese men had a significantly higher risk of death for all causes and for cancer. Cardiovascular risk factors may act as effect modifiers in these associations

Single-cell Atlas of common variable immunodeficiency shows germinal center-associated epigenetic dysregulation in B-cell responses

Common variable immunodeficiency (CVID), the most prevalent symptomatic primary immunodeficiency, displays impaired terminal B-cell differentiation and defective antibody responses. Incomplete genetic penetrance and ample phenotypic expressivity in CVID suggest the participation of additional pathogenic mechanisms. Monozygotic (MZ) twins discordant for CVID are uniquely valuable for studying the contribution of epigenetics to the disease. Here, we generate a single-cell epigenomics and transcriptomics census of naïve-to-memory B cell differentiation in a CVID-discordant MZ twin pair. Our analysis identifies DNA methylation, chromatin accessibility and transcriptional defects in memory B-cells mirroring defective cell-cell communication upon activation. These findings are validated in a cohort of CVID patients and healthy donors. Our findings provide a comprehensive multi-omics map of alterations in naïve-to-memory B-cell transition in CVID and indicate links between the epigenome and immune cell cross-talk. Our resource, publicly available at the Human Cell Atlas, gives insight into future diagnosis and treatments of CVID patients