Screening of Synergic Interactions of Anti-Angiogenic and Anti-Tumor Compounds

Solid cancers have several common characteristics that Hanahan & Weinberg named as the hallmarks of cancer. Angiogenesis is an essential hallmark of cancer because tumor cells need oxygen and nutrients delivered by the vascular system. In fact, tumor growth and metastasis are angiogenesis dependent, and microvascular endothelial cells recruited by tumors have become an important target in cancer therapy. Combinations of drugs with different modes of action may lead to enhanced antitumor and antiangiogenic effects without injuring the host. The combined use of two drugs may sometimes produce enhanced, unchanged or diminished effects in comparison with their individual effects. These three different types of behaviour of the interacting drugs are called synergy, additive/indifferent and antagonistic effects. In the present work, we analyze 105 paired combinations of 15 compounds, some described by our research group as potent antiangiogenic compounds, and others currently used in clinical therapy. Our results show synergistic effects of several paired combinations using the MTT assay. [Our experimental work is supported by grants BIO2014-56092-R (MINECO and FEDER), P12-CTS-1507 (Andalusian Government and FEDER) and funds from group BIO-267 (Andalusian Government). The "CIBER de Enfermedades Raras" is an initiative from the ISCIII (Spain)].This communication has the support of a travel grant "Universidad de Málaga.Campus de Excelencia Internacional Andalucía Tech".Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Del revés: metaloproteinasa de la matriz extracelular 2 y su función en el nucleolo

Las metaloproteinasas de la matriz extracelular (MMPs) son proteínas que tradicionalmente han sido ubicadas en el exterior celular y su función ha sido relacionada con la remodelación de la matriz extracelular en procesos tan diversos como el desarrollo embrionario, durante la reparación de daños tisulares y en patologías tumorales, en concreto en distintos tipos de cáncer sólido. Aunque diversas MMPs han sido halladas en el interior celular inclusive en compartimentos subcelulares específicos donde desempeñan funciones concretas. La metaloproteinasa de la matriz extracelular 2 (MMP-2) ha sido localizada en el interior de diversos tipos celulares como cardiomiocitos de rata y ratón, linfocitos y en distintos tipos de células tumorales. Además de la localización citoplasmática de MMP-2, esta también ha sido encontrada en mitocondrias, en membranas asociadas a la mitocondria, así como en el núcleo celular. Aunque la actividad de MMP-2 mitocondrial ha sido estudiada en cardiomiocitos bajo condiciones de isquemia y reperfusión, y su activación está directamente relacionada con el incremento del estrés oxidativo, sin embargo, el conocimiento acerca de MMP-2 nuclear es escaso. El interés por averiguar el papel de MMP-2 en el núcleo celular ha representado un reto por la complejidad que conlleva la biología del núcleo y por la inesperada localización de esta proteína. Nuestro interés por dilucidar el papel de MMP-2 en el núcleo celular, nos ha llevado a identificar que MMP-2 se encuentra enriquecida en cierta sublocalización nuclear como son los nucleolos, donde MMP-2 está relacionada con la transcripción del DNA ribosomal.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Hydroxytyrosol targets extracellular matrix remodeling by endothelial cells and inhibits both ex vivo and in vivo angiogenesis

This is the preprint version of our manuscript, corresponding to the article that has been 
published in final form at FOOD CHEMISTRY with DOI: 10.1016/j.foodchem.2016.10.111The health benefits of olive oil are attributed to their bioactive compounds, such as hydroxytyrosol. Previously, we demonstrated that hydroxytyrosol inhibits angiogenesis in vitro. The present study aimed to: i) get further insight into the effects of hydroxytyrosol on extracellular matrix remodeling; and ii) test whether hydroxytyrosol is able to inhibit angiogenesis ex vivo and in vivo. Hydroxytyrosol induced a shift toward inhibition of proteolysis in endothelial cells, with decreased expression of extracellular matrix remodeling-enzyme coding genes and increased levels of some of their inhibitors. Furthermore, this work demonstrated that hydroxytyrosol, at concentrations within the range of its content in virgin olive oil that can be absorbed from moderate and sustained virgin olive oil consumption, is a strong inhibitor of angiogenesis ex vivo and in vivo. These results suggest the need for translational studies to evaluate the potential use of hydroxytyrosol for angio-prevention and angiogenesis inhibition in clinical setting.This work was supported by grants BIO2014-56092-R (MINECO and FEDER) and P12-CTS-1507 (Andalusian Government and FEDER). The “CIBER de Enfermedades Raras” is an initiative from the ISCIII (Spain). The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript

Estratigrafía Sísmica de Alta Resolución en las Rías Baixas: Pontevedra y Vigo (NO España)

El estudio detallado de los perfiles de sísmica reflexión de alta resolución, de las rías de Pontevedra y Vigo ha permitido diferenciar 7 unidades sísmicas. Las unidades sísmicas Bg y Bm se interpretan como correspondientes a rocas del basamento, graníticas y metamórficas respectivamente. Las 5 unidades sísmicas restantes corresponden al relleno sedimentario de las rías. La identificación de los límites de secuencia L2 y L3 permiten reconocer tres secuencias sedimentarias (S1, S2 y S3), de más antigua a más reciente, que van marcando una progresiva ampliación de la zona de sedimentación en las rías. En la secuencia más reciente (S3) destaca la presencia de apantallamientos acústicos y/o formas de escape, interpretadas como debidas a la presencia de gas biogénico en los sedimentos.Detailed study of high-resolution seismic reflection profiles of the Ría de Pontevedra and Vigo (NW Spain) has allowed to distinguish 7 seismic units. Seismic units Bg and Bm are interpreted as granitic and methamorphic basement rocks respectively. The other 5 seismic units correspond to the sedimentary infill of the rías. The identification of the sequences boundaries L2 and L3 allow to recognize three sedimentary sequences (S1, S2 and S3), from oldest to youngest, 217 S. García-Gil et al. Estratigrafía sísmica de alta resolución en las Rías Baixas:... that give a progressive enlargement of the rías sedimentary basin. In the most recent sequence (S3) appear acoustic blanking and/or escape features, interpreted as due to the presence of biogenic gas in the sediments of the rías

The noni anthraquinone damnacanthal is a multi-kinase inhibitor with potent anti-angiogenic effects

Este es el manuscrito que fue aceptado y que finalmente se publicó en Cancer Letters con el DOI: 10.1016/j.canlet.2016.10.037The natural bioactive compound damnacanthal inhibits several tyrosine kinases. Herein, we show that -in fact- damancanthal is a multi kinase inhibitor. A docking and molecular dynamics simulation approach allows getting further insight on the inhibitory effect of damnacanthal on three different kinases: vascular endothelial growth factor receptor-2, c-Met and focal adhesion kinase. Several of the kinases targeted and inhibited by damnacanthal are involved in angiogenesis. Ex vivo and in vivo experiments clearly demonstrate that, indeed, damnacanthal is a very potent inhibitor of angiogenesis. A number of in vitro assays contribute to determine the specific effects of damnacanthal on each of the steps of the angiogenic process, including inhibition of tubulogenesis, endothelial cell proliferation, survival, migration and production of extracellular matrix remodeling enzyme. Taken altogether, these results suggest that damancanthal could have potential interest for the treatment of cancer and other angiogenesisdependent diseases.Supported by grants BIO2014-56092-R (MINECO and FEDER), P12-CTS-1507 (Andalusian Government and FEDER) and funds from group BIO-267 (Andalusian Government). The "CIBER de Enfermedades Raras" is an initiative from the ISCIII (Spain). JAGV had the financial support of Vicerrectorado de Investigación y Transferencia (University of Málaga, Spain). The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript

Screening of Synergistic Interactions of Epigallocatechin-3-gallate with Antiangiogenic and Antitumor Compounds

This is the accepted manuscript of the article that finally was published in Synergy with DOI: 10.1016/j.synres.2016.05.001Purpose: To screen for possible synergistic interactions of epigallocatechin-3-gallate (EGCG) with a selection of 10 anti-angiogenic or anti-tumor compounds on the survival of endothelial and tumor cells. Methods: Human HMEC endothelial and MDA-MB231 breast cancer cells were treated with different concentrations of EGCG and the 10 tested compounds either as single agents or in paired combinations with EGCG for 3 days and final survival of cells was determined by the MTT assay. IC50 values, sensitization factors and combination indexes were calculated. Results; IC50 values of 140±2 and 45±6 μM were determined for EGCG-treated endothelial and tumor cells, respectively. IC50 values for all tested compounds were within the micromolar and the submillimolar range. The values of the sensitization factor increased and those of the combination index decreased for paired combinations of EGCG with 4-methylumbelliferone. The opposite was true for the combination of EGCG with vitamin D3. Other tested combinations did not exhibit a clear monotonic effect but rather a biphasic behaviour. Conclusion: Combinations of EGCG and 4-methylumbelliferone synergistically decrease endothelial and tumor cell survival. In contrast, the presence of EGCG antagonizes with the antiproliferative effect exerted by vitamin D3 on endothelial and tumor cells.Supported by grants BIO2014-56092-R (MINECO and FEDER), P12-CTS-1507 (Andalusian Government and FEDER) and funds from group BIO-267 (Andalusian Government). The "CIBER de Enfermedades Raras" is an initiative from the ISCIII (Spain). JAGV has the financial support of Vicerrectorado de Investigación y Transferencia (University of Málaga, Spain). The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript

Design of Carvacrol-based active packaging for extending fresh fish shelf-life

Shelf life is the time-span where the product is in good conditions for consumption, from either safety or quality points of view. Shelf life is defined to guarantee food safety (“use-by” date), or to guarantee both safety and quality food standards (“best-before date”). The aim of active packaging systems is to extend shelf-life by releasing active substances into the food product. The active packaging is composed of one or several layers. Each layer may contain a given concentration of the active substance whose release velocity will depend on the layer material. An adequate selection of the layer composition and active substance initial concentration will have an impact on shelf life. In this work, we use mathematical models in combination with optimization methods to (i) design the optimal configuration of the smart packaging for both “use-by date” and “best-before date” criteria; and (ii) assess and predict shelf life changes according to variations on storage conditions. Two different types of models are used: (i) release of the active substance into the food product [1]; and (ii) shelf-life evolution as a function of storage conditions and active substance concentration in the food [2]. The growth of Listeria monocytogenes, which affects widely consumed products, is used as the safety indicator. The KI-value, which is related to ATP-degradation compounds, is used as the quality indicator. Carvacrol is a substance that inhibits bacterial growth. In this work, it will be used as the active substance to limit the growth of Listeria monocytogenes (food safety) and other spoilage bacteria responsible for quality changes. The optimal design allows for an increase in shelf-life of around 24% as compared with the fish product without active packaging

Aplicación del Sonar de Barrido Lateral a la cartografía de fondos marinos. Ejemplo de la Ría de Pontevedra (Galicia, NO España).

Se realiza una cartografía de detalle de los fondos actuales de la Ría de Pontevedra basándose en los registros de sonar de barrido lateral. El análisis de los sonogramas obtenidos permite diferenciar cuatro patrones de reflectividad. Los datos de muestras directas del fondo obtenidas dentro de la zona que caracteriza a cada patrón establecen la relación directa entre reflectividad-textura. La correlación de las sonografías con la geología de la zona emergida hacen posible distinguir en los registros los dos tipos de basamento (granítico y metamórfico) en función de sus reflectividades y rasgos estructurales (fracturas). El sistema acústico aplicado permite discriminar entre formas sedimentarias (megaripples) y antrópicas (bateas, marcas de arrastre), a la vez que se establece su relación directa con las condiciones hidrodinámicas y rasgos morfológicos de la Ría de Pontevedra.Side scan sonar is used to provide a high resolution mapping of the seafloor of the Ría de Pontevedra. Sonograph analysis allows us to distinguish four backscatter patterns. Bottom samples within each pattern establish a direct relationship between backscatter and sediment texture. Sonograph correlation with land geology permit the differentiation between granitic and metamorphic basement, as well as its structural features (fractures). The side scan sonar is also used for discriminating sedimentary (megaripples) and anthropogenic features (mussel rafts, trawl-marks) and also for establishing the relationship between the hydrodynamic conditions and geomorphology of the Ría

Model-based design of smart active packaging systems with antimicrobial activity

Smart active packaging is an innovative packaging system that combines the benefits of measuring, estimating or predicting different aspects of food quality or safety with the release of an active substance that extends product shelf life. Nevertheless, in its typical configuration, the active packaging and the smart packaging are not connected, and the information provided is not exploited to design the release of the active substance. In this work, we demonstrate how smart active packaging systems using predictive mathematical models allow the automatic optimisation of food packaging design and the prediction of the expected shelf life along the food chain. On the one hand, the system calculates the best design of the active packaging and the concentration of the active substance in the different layers that maximise food quality and safety. On the other hand, the model allows to calculate and update shelf life values along the food chain under unexpected changes in the storage conditions. Shelf life estimations and prediction will help distributors and sellers to adjust the product market prices. For example, prices can be lowered to avoid food losses when the product is close to its use-by date. Hake (Merluccius merluccius) represents an example of a highly relevant and perishable food that can be conserved using natural antimicrobials. Therefore, the case study selected to illustrate the proposed methodology consists of the smart active packaging of hake using carvacrol as the active substance (antimicrobial). Besides, different polymers are considered as possible active packaging materials. The Matlab™ codes required to perform the simulations of the models described in this work as well as the optimisations for packaging design are available at https://doi.org/10.5281/zenodo.3244153C.V. acknowledges funding received from the European Union's Horizon 2020 research and innovation programme under grant agreement no. 723575 (CoPro project). M.M.I. belongs to the Galician Competitive Research Group (ED431C 2017/029) and the CRETUS strategic partnership (AGRUP2017/01), co-funded by FEDER (EU). M.R.G. acknowledges financial support through the projects IMMICRO (201870E134) and ControlAR (RTI2018-093560-J-I00, MCIU/AEI/FEDER, UE)S

Análisis de compuestos naturales candidatos a fármacos por su actividad antitumoral, antiinflamatorio o antiangiogénico

El término cáncer, enfermedad neoplásica o enfermedad tumoral, integra a una gran diversidad de condiciones patológicas con muy distintas etiologías, desarrollo y pronóstico. En la evolución de un tumor o progresión tumoral pueden distinguirse tres fases: 1. Transformación neoplásica. 2. Progresión tumoral in situ dependiente de angiogénesis. 3. Invasión y metástasis. Cada una de estas etapas supone la acumulación de “fallos” en los diversos y sofisticados sistemas de regulación del organismo. Estos “fallos” consisten básicamente en la acumulación de mutaciones en genes cuyos productos juegan un importante papel en dichos sistemas de regulación. No todas las enfermedades neoplásicas pasan por estas tres fases. Así, en las leucemias no puede hablarse de un crecimiento in situ al tratarse de células sanguíneas circulantes. Por otra parte, hay numerosos tumores que permanecen encapsulados y nunca manifiestan capacidad invasiva y por tanto no dan lugar a metástasis. En la mayoría de los casos la carcinogénesis (etapa de transformación neoplásica) no es el principal problema del cáncer desde un punto de vista clínico, puesto que en la mayoría de los casos del cáncer humano las muertes son realmente causadas por las metástasis. El cáncer se ha convertido en la primera causa de mortandad en los países industrializados. Por ello, en las cuatro últimas décadas se ha investigado mucho acerca de la biología del cáncer. Fruto de ello, se ha generado conocimiento sobre los factores que favorecen el desarrollo del cáncer y los mecanismos moleculares que dan lugar a la transformación en células cancerígenas. Ello se ha utilizado para desarrollar estrategias terapéuticas, en primer lugar por la diferencia en la tasa de replicación de las células cancerígenas en comparación a las células sanas, y posteriormente, atendiendo a los mecanismos moleculares que caracterizan a las células cancerígenas. La identificación de las así denominadas “señales distintivas del cáncer” ha abierto una nueva vía de exploración de terapias dirigidas. La presente Tesis Doctoral se enmarca dentro de la principal línea de investigación de nuestro grupo, centrada en la identificación y caracterización de nuevos compuestos antitumorales, antiinflamatorios o antiangiogénicos. El grupo de investigación en el que se ha realizado la presente Tesis Doctoral tiene un extenso historial previo en oncología básica en general y en la identificación de compuestos antiangiogénicos y antiinflamatorios en particular. De acuerdo con los intereses de la línea de investigación del grupo, la presente Tesis Doctoral nace con el propósito de buscar y caracterizar nuevos compuestos naturales con potencial antitumoral, antiinflamatorio o antiangiogénico. Se han realizado ensayos in vitro, ex vivo e in vivo para la caracterización de los compuestos seleccionados, contribuyendo al conocimiento de su modo de acción. Además, se ha realizado una primera búsqueda sistemática de efectos sinérgicos en combinaciones pareadas de compuestos antitumorales o antiangiogénicos con distintos mecanismos de acción. Esta Tesis Doctoral identifica dos nuevo compuestos naturales antiangiogénicos y describe sus efectos in vitro, ex vivo e in vivo. Además, esta Tesis Doctoral identifica uno de estos dos compuestos como un potente inhibidor de tirosina cinasas que, a través de su efecto inhibidor sobre c-Met, se perfila como un nuevo compuesto prometedor para el tratamiento de cáncer de hígado. En esta Tesis Doctoral también hemos ampliado el espectro de acciones biológicas atribuibles al compuesto natural aeroplisinina-1 (previamente descrito como anti-angiogénico por nuestro grupo), identificándolo como inhibidor de moléculas pro-inflamatorias. Finalmente, esta Tesis Doctoral abre un nuevo camino de investigación en el grupo para la búsqueda de nuevas estrategias antitumorales, antiinflamatorias o antiangiogénicas basado en el efecto que ejercen combinaciones de fármacos sobre las etapas claves en los proceso tumoral y angiogénico