c-Src signaling in triple negative breast cancer cells: role of Cyr61

Póster presentado al CNIO Frontiers Meeting: "Metastasis Initiation: Mechanistic Insights and Therapeutic Opportunities", celebrado en el Spanish National Cancer Research Centre (CNIO) Auditorium del 28 al 30 de septiembre de 2015.The SFKs (Src Family Kinases) control cellular pathways involved in division, motility, adhesion, angiogenesis, and survival. Therefore, their deregulation is associated with tumorigenesis, and metastasis. c-Src is overexpressed and/or aberrantly activated in epithelial tumors: pancreatic, colorectal, prostatic, ovarian, breast, etc. We previously showed that SFKs catalytic inhibitors (Dasatinib, PP2, and SU6656) reduce proliferation, migration, and invasiveness of MDA-MB-231. Here, we analyzed c-Src contribution to initial steps of metastasis by Tet-On conditional expression of a specific shRNA-c-Src, which suppressed c-Src mRNA and protein levels in MDA-MB-231. c-Src suppression did not alter cell proliferation or survival, but it significantly reduced anchorage-independent growth. Concomitantly with diminished tyrosine-phosphorylation/activation of Fak, caveolin-1, paxillin and p130CAS, c-Src depletion inhibited migration, invasion, transendothelial migration, and reduced MMP2, MMP7 and MMP9 in secretome. Quantitative proteomic analyses of secretome showed that Cyr61 levels, detected in exosomal fraction, were diminished upon shRNA-c-Src expression. However, Cyr61 expression was unaltered inside cells. Cyr61 partially colocalized with cis-Golgi gp74 marker, and with exosomal marker CD63, but c-Src depletion did not alter their distribution. In SUM159PT, transient c-Src suppression also reduced secreted exosomal Cyr61. Furthermore, conditional expression of c-Src dominant negative mutant (c-Src-K295M/Y527F) in MDA-MB-231 and in SUM159PT diminished secreted Cyr61 as well. Cyr61 transient suppression in MDA-MB-231 inhibited invasion and transendothelial migration. Finally, in both MDA-MB-231 and SUM159PT, a neutralizing Cyr61 antibody restrained migration. Collectively, these results suggest that c-Src regulates secreted proteins, including exosomal Cyr61, which are involved in modulating the metastatic potential of triple negative breast cancer cells.Peer Reviewe

Cobalt oxide-Carbon nanocatalysts with highly enhanced catalytic performance for the green synthesis of nitrogen heterocycles through Friedländer condensation.

A novel series of eco-sustainable catalysts developed by supporting CoO nanoparticles on different carbon supports, highly efficient in the synthesis of quinolines and naphthyridines, through the Friedländer condensation, are reported for the first time. Textural properties, dispersion and location of the Co-phase are influenced by the nature of the carbon support, Co-precursor salt and metal loading, having a significant impact on the catalytic performance. Thus, the presence of the mesopores and macropores in carbon aerogels together with the homogeneous distribution of the active phase favours the formation of product 3a as a function of the metal loading. However, an increase in the metal content when using CNTs indicates the formation of CoO aggregates and an optimal concentration of 3 wt% CoO was observed, providing the highest conversion values. The carbon-based catalysts herein reported can be considered to be a sustainable alternative having advantages such as easy preparation, superior stability and notably enhanced catalytic performance, operating at lower temperature and under solvent-free conditions.pre-print815 K

Synthesis of large-pore zeolites from chiral structure-directing agents with two l-prolinol units

In this work, we perform an in-depth experimental and computational study about the structure-directing effect of two new chiral organic quaternary ammonium dications bearing two N-methyl-prolinol units linked by a xylene spacer in para or meta relative orientation, displaying four enantiopure stereogenic centers in (S) configuration. Synthesis results show that the para-xylene derivative is an efficient structure-directing agent, promoting the crystallization of ZSM-12 (in pure-silica composition), beta zeolite (as pure-silica, or in the presence of Al or Ge), and a mixture of polymorphs C, A and B of zeolite beta (in the presence of Ge). In contrast, the meta-xylene derivative showed a much poorer structure-directing activity, yielding only amorphous materials unless Ge is present in the gel, where beta and polymorph C (together with A and B) zeolites crystallized. Molecular simulations showed that the para-xylene dication displays a cylindrical shape suitable for confining in zeolite pores, while the meta-xylene derivative has an angular shape that shifts from the typical dimensions required for 12MR zeolite channels. Despite enantio-purity of the para-xylene dication with (S, S, S, S) configuration, no enrichment in polymorph A of the zeolite beta samples obtained was observed by Transmission Electron Microscopy. With the aid of molecular simulations, the failure in transferring chirality to the zeolite is explained by the loose fit of this SDA in the large-pores of zeolite beta, and a lack of close geometrical fit with the chiral element of polymorph A, as evidenced by the very similar interaction of the cation with the two enantiomorphic space groups of polymorph A. Nevertheless, the molecular-level knowledge gained in this work can provide insights for the future design of more efficient SDAs towards the synthesis of chiral zeolites

Structural analysis of IPC zeolites and related materials using positron annihilation spectroscopy and high-resolution argon adsorption

ETH authors thanks for the grant ETH 33 15-1. PE and JČ acknowledge the financial support from the Czech Science Foundation (P106/12/0189). JPR and JČ gratefully acknowledge the financial support from the European Union Seventh Framework Programme (FP7/ 2007-2013) under grant agreement no. 604307. HRTEM characterization was performed at the Advanced Microscopy Laboratory (LMA) and the research leading to these results has received funding from the European Union Seventh Framework Programme under Grant Agreement 312483 – ESTEEM2 (Integrated Infrastructure Initiative-I3).The advanced investigation of pore networks in isoreticular zeolites and mesoporous materials related to the IPC family was performed using high-resolution argon adsorption experiments coupled with the development of a state-of-the-art non-local density functional theory approach. The optimization of a kernel for model sorption isotherms for materials possessing the same layer structure, differing only in the interlayer connectivity (e.g. oxygen bridges, single- or double-four-ring building units, mesoscale pillars etc.) revealed remarkable differences in their porous systems. Using high-resolution adsorption data, the bimodal pore size distribution consistent with crystallographic data for IPC-6, IPC-7 and UTL samples is shown for the first time. A dynamic assessment by positron annihilation lifetime spectroscopy (PALS) provided complementary insights, simply distinguishing the enhanced accessibility of the pore network in samples incorporating mesoscale pillars and revealing the presence of a certain fraction of micropores undetected by gas sorption. Nonetheless, subtle differences in the pore size could not be discriminated based on the widely-applied Tao-Eldrup model. The combination of both methods can be useful for the advanced characterization of microporous, mesoporous and hierarchical materials.PostprintPeer reviewe

GTM-3, an extra-large pore enantioselective chiral zeolitic catalyst

The development of chiral zeolitic catalysts possessing extra-large pores and endowed with the capability of enantioselectively processing bulky products represents one of the greatest challenges in chemistry. Here, we report the discovery of GTM-3, an enantio-enriched extra-large pore chiral zeolite material with -ITV framework structure, obtained using a simple enantiopure organic cation derived from the chiral pool, N,N-ethyl-methyl-pseudoephedrinium, as the chiral-inductor agent. We demonstrate the enantio-enrichment of GTM-3 in one of the two enantiomorphic polymorphs using the two enantiomers of the organic cation. Interestingly, we prove the ability of this zeolitic material to perform enantioselective catalytic operations with very large substrates, here exemplified by the catalytic epoxide aperture of the bulky trans-stilbene oxide with alcohols, yielding unprecedented product enantiomeric excesses up to 30%. Our discovery opens the way for the use of accessible chiral zeolitic materials for the catalytic asymmetric synthesis of chiral pharmaceutical compounds

Nanoalloying in real time: a high resolution STEM and computer simulation study

Bimetallic nanoparticles constitute a promising type of catalysts, mainly because their physical and chemical properties may be tuned by varying their chemical composition, atomic ordering, and size. Today, the design of novel nanocatalysts is possible through a combination of virtual lab simulations on massive parallel computing and modern electron microscopy with picometre resolution on one hand, and the capability of chemical analysis at the atomic scale on the other. In this work we show how the combination of theoretical calculations and characterization can solve some of the paradoxes reported about nanocatalysts: Au-Pd bimetallic nanoparticles. In particular, we demonstrate the key role played by adsorbates, such as carbon monoxide (CO), on the structure of nanoalloys. Our results imply that surface condition of nanoparticles during synthesis is a parameter of paramount importance.Fil: Mariscal, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; ArgentinaFil: Mayoral, Alba. Universidad de Zaragoza. Instituto de Nanociencia de Aragón; EspañaFil: Olmos Asar, Jimena Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; ArgentinaFil: Magen, César. Universidad de Zaragoza. Instituto de Nanociencia de Aragón; EspañaFil: Mejia Rosales, Sergio Javier. Universidad Autónoma de Nuevo León; MéxicoFil: Pérez Tijerina, Eduardo. Universidad Autónoma de Nuevo León; MéxicoFil: José Yacamán, Miguel. University of Texas; Estados Unido

c-Src functionality controls self-renewal and glucose metabolism in MCF7 breast cancer stem cells

Deregulation of Src kinases is associated with cancer. We previously showed that SrcDN conditional expression in MCF7 cells reduces tumorigenesis and causes tumor regression in mice. However, it remained unclear whether SrcDN affected breast cancer stem cell functionality or it reduced tumor mass. Here, we address this question by isolating an enriched population of Breast Cancer Stem Cells (BCSCs) from MCF7 cells with inducible expression of SrcDN. Induction of SrcDN inhibited self-renewal, and stem-cell marker expression (Nanog, Oct3-4, ALDH1, CD44). Quantitative proteomic analyses of mammospheres from MCF7-Tet-On-SrcDN cells (data are available via ProteomeXchange with identifier PXD017789, project DOI: 10.6019/PXD017789) and subsequent GSEA showed that SrcDN expression inhibited glycolysis. Indeed, induction of SrcDN inhibited expression and activity of hexokinase, pyruvate kinase and lactate dehydrogenase, resulting in diminished glucose consumption and lactate production, which restricted Warburg effect. Thus, c-Src functionality is important for breast cancer stem cell maintenance and renewal, and stem cell transcription factor expression, effects linked to glucose metabolism reduction.This work has been supported by grand SAF2016–75991-R (MINECO, AEI/FEDER, UE) to Jorge Martín-Pérez and ISCIII [grand PI 16/00789] to Miguel Ángel Fernández-Moreno. Víctor Mayoral-Varo was supported by the grand SAF2016–75991-R (MINECO, AEI/FEDER, UE). We acknowledge support for publication fee by the CSIC Open Access Publication Support Initiative through its Unit for Information Resources for Research (URICI)

Monitoring new long-lasting intravitreal formulation for glaucoma with vitreous images using optical coherence tomography

Intravitreal injection is the gold standard therapeutic option for posterior segment patholo-gies, and long-lasting release is necessary to avoid reinjections. There is no effective intravitreal treatment for glaucoma or other optic neuropathies in daily practice, nor is there a non-invasive method to monitor drug levels in the vitreous. Here we show that a glaucoma treatment combining a hypotensive and neuroprotective intravitreal formulation (IF) of brimonidine–Laponite (BRI/LAP) can be monitored non-invasively using vitreoretinal interface imaging captured with optical coherence tomography (OCT) over 24 weeks of follow-up. Qualitative and quantitative characterisation was achieved by analysing the changes in vitreous (VIT) signal intensity, expressed as a ratio of retinal pigment epithelium (RPE) intensity. Vitreous hyperreflective aggregates mixed in the vitreous and tended to settle on the retinal surface. Relative intensity and aggregate size progressively decreased over 24 weeks in treated rat eyes as the BRI/LAP IF degraded. VIT/RPE relative intensity and total aggregate area correlated with brimonidine levels measured in the eye. The OCT-derived VIT/RPE relative intensity may be a useful and objective marker for non-invasive monitoring of BRI/LAP IF

An internet-based intervention for depression in primary care in Spain: A randomized controlled trial

Background: Depression is the most prevalent cause of illness-induced disability worldwide. Face-to-face psychotherapeutic interventions for depression can be challenging, so there is a need for other alternatives that allow these interventions to be offered. One feasible alternative is Internet-based psychological interventions. This is the first randomized controlled trial (RCT) on the effectiveness of an Internet-based intervention on depression in primary health care in Spain. Objective: Our aim was to compare the effectiveness of a low-intensity therapist-guided (LITG) Internet-based program and a completely self-guided (CSG) Internet-based program with improved treatment as usual (iTAU) care for depression. Methods: Multicenter, three-arm, parallel, RCT design, carried out between November 2012 and January 2014, with a follow-up of 15 months. In total, 296 adults from primary care settings in four Spanish regions, with mild or moderate major depression, were randomized to LITG (n=96), CSG (n=98), or iTAU (n=102). Research completers at follow-up were 63.5%. The intervention was Smiling is Fun, an Internet program based on cognitive behavioral therapy. All patients received iTAU by their general practitioners. Moreover, LITG received Smiling is Fun and the possibility of psychotherapeutic support on request by email, whereas CSG received only Smiling is Fun. The main outcome was the Beck Depression Inventory-II at 3 months from baseline. Mixed-effects multilevel analysis for repeated measures were undertaken. Results: There was no benefit for either CSG (B coefficient=-1.15; P=.444)] or LITG (B=-0.71; P=.634)] compared to iTAU, at 3 months. There were differences at 6 months iTAU vs CSG (B=-4.22; P=.007); iTAU vs LITG (B=-4.34; P=.005)] and 15 months iTAU vs CSG (B=-5.10; P=.001); iTAU vs LITG (B=-4.62; P=.002)]. There were no differences between CSG and LITG at any time. Adjusted and intention-to-treat models confirmed these findings. Conclusions: An Internet-based intervention for depression combined with iTAU conferred a benefit over iTAU alone in the Spanish primary health care system

Predictors of Adherence in Three Low-Intensity Intervention Programs Applied by ICTs for Depression in Primary Care

Depression is one of the most common disorders in psychiatric and primary care settings, and is associated with disability, loss in quality of life, and economic costs. Internet-based psychological interventions have been shown to be effective in depression treatment but present problems with a low degree of adherence. The main aim of this study is to analyze the adherence predictors in three low-intensity interventions programs applied by Information and Communication Technologies (ICTs) for depression. A multi-center, randomized, controlled clinical trial was conducted with 164 participants with depression, who were allocated to: Healthy Lifestyle Program, Positive Affect Promotion Program or Mindfulness Program. Sociodemographic characteristics, Patient Health Questionnaire-9, Visual Analog Scale, Short Form Health Survey, Positive and Negative Affect Schedule, Five Facets Mindfulness Questionnaire, Pemberton Happiness Index and Treatment Expectancy Questionnaire were used to study adherence. Results showed that positive affect resulted in a predictor variable for Healthy Lifestyle Program and Positive Affect Promotion Program. Perceived health was also a negative adherence predictor for the Positive Affect Promotion Program. Our findings demonstrate that there are differences in clinical variables between treatment completers and non-completers and we provide adherence predictors in two intervention groups. Although new additional predictors have been examined, further research is essential in order to improve tailored interventions and increase adherence treatment