Search

Fundamental biological processes of cell identity and cell fate determination are controlled by complex regulatory networks. These processes require molecular mechanisms that confer cellular phenotypic memory and state persistence. In this minireview, we will summarize mechanisms of cell memory based on regulatory hysteretic feedback loops and explore epigenetic mechanisms widely represented in nature, with special focus on epithelial-to-mesenchymal plasticity. We will also discuss the functional consequences of cell memory and epithelial-to-mesenchymal plasticity dynamics during development and cancer metastasis.

Background: The role of MET alterations in non-small cell lung cancer (NSCLC) is increasing and several targeted agents are under evaluation. MET exon 14 skipping mutations and MET amplifications are associated with potential sensitivity to MET inhibition, though resistance mechanisms are emerging. In MET addicted cells, MET inhibition leads to activation of proviral integration site for Moloney murine leukemia virus-1 (PIM1). PIM1 and proto-oncogene tyrosine-protein kinase Src (SRC) can regulate the expression of receptor tyrosine kinases (RTKs), potentially inducing resistance to MET inhibition through cross-activation. Methods: We evaluated the activity of class I-II MET inhibitors, the SRC inhibitor dasatinib, and panPIM inhibitors in four MET addicted cell lines. We assessed the effect of the dual MET/PIM and MET/ SRC inhibition on cell viability and at the protein level. We evaluated RNA expression profiles of the cell lines. Advanced NSCLCs were also screened for MET alterations. Results: All cell lines were sensitive to class I-II MET inhibitors. All cell lines were resistant to single PIM and SRC inhibition. Dual MET/PIM inhibition was synergistic or additive in MET amplified cell lines and dual MET/SRC inhibition was highly synergistic in all MET addicted cell lines. The addition of an SRC inhibitor partially prevents the RTKs cross-activation. MET alterations were found in 9 out of 97 evaluable samples (9.3%); median overall survival in MET altered patients was 5 months (95% CI, 3 m-NA). Conclusions: We identified a potential role of PIM inhibition in MET amplified tumors and of SRC inhibition in MET addicted tumors. Potential applications of this new treatment strategy warrant further evaluation.

Introduction Epidermal growth factor receptor (EGFR) pathway deregulation promotes the acquisition of stemlike properties in non–small-cell lung cancer. EGFR inhibition through NOTCH enriches lung cancer stem cells (CSCs). Src through Yes-associated protein 1 (YAP1) activates NOTCH. Signal transduction and activator of transcription 3 (STAT3) activation occurs upon EGFR blockade and regulates the generation of CSCs. Patients and Methods Using the Aldefluor assay kit, we investigated the enrichment of aldehyde dehydrogenase (ALDH)-positive cells in EGFR-mutation–positive cells treated with gefitinib, afatinib, and osimertinib. Western blot analysis was performed to evaluate changes in CSC marker expression upon EGFR blockade. We performed gene expression analysis in a cohort of EGFR-mutation–positive non–small-cell lung cancer patients. We evaluated the association of gene expression with treatment outcomes. Results The cell subpopulation surviving EGFR inhibition had high ALDH activity and elevated CSC marker expression. Concurrent inhibition of EGFR, STAT3, and Src diminished the CSC subpopulation in an EGFR-mutation–positive cellular model. In a cohort of 64 EGFR-mutation–positive patients, 2 ALDH1 isoforms and the NOTCH target hairy and enhancer of split 1 (HES1), when highly expressed, were predictive of worse outcome to EGFR blockade. The gene expression of B-cell–specific Moloney murine leukemia virus integration site 1 (Bmi-1) that maintains the self-renewal of stem cells was also related to treatment outcome. Conclusion Single EGFR inhibitors increase the population of CSCs. Combinatory therapy targeting STAT3 and Src may be of potential benefit. ALDH1, HES1, and Bmi-1 are essential biomarkers in the initial assessment of EGFR-mutation–positive patients.

Ligand-dependent corepressor (LCOR) mediates normal and malignant breast stem cell differentiation. Cancer stem cells (CSCs) generate phenotypic heterogeneity and drive therapy resistance, yet their role in immunotherapy is poorly understood. Here we show that immune-checkpoint blockade (ICB) therapy selects for LCORlow CSCs with reduced antigen processing/presentation machinery (APM) driving immune escape and ICB resistance in triple-negative breast cancer (TNBC). We unveil an unexpected function of LCOR as a master transcriptional activator of APM genes binding to IFN-stimulated response elements (ISREs) in an IFN signaling-independent manner. Through genetic modification of LCOR expression, we demonstrate its central role in modulation of tumor immunogenicity and ICB responsiveness. In TNBC, LCOR associates with ICB clinical response. Importantly, extracellular vesicle (EV) Lcor-messenger RNA therapy in combination with anti-PD-L1 overcame resistance and eradicated breast cancer metastasis in preclinical models. Collectively, these data support LCOR as a promising target for enhancement of ICB efficacy in TNBC, by boosting of tumor APM independently of IFN. This work was supported by the Instituto de Salud Carlos III-FSE (nos. MS17/00037 and PI18/00014) and the Cancer Research Institute, Clinic and Laboratory Integration Program (grant no. CRI2477 to T.C.-T). We also thank the AECC LAB (grant no. LABAE19007CELI) and the FERO foundation (to T.C.-T). This work was also supported by ISCIII (CIBERONC nos. CB16/12/00481, CB16/12/00241, PI18/00006 and PI21/00002), Generalitat de Catalunya (no. 2017 SGR 507) and the European Community through the Regional Development Funding Program to J. Albanell. Y.K. is supported by the Brewster Foundation, the Breast Cancer Research Foundation, the Susan G. Komen Foundation and the American Cancer Society. J. Arribas is funded by the Breast Cancer Research Foundation (no. BCRF-20-08), Instituto de Salud Carlos III (no. PI19/01181), Asociación Española Contra el Cáncer (no. GCAEC19017ARRI) and Fundación BBVA (no. CAIMI VHIO-FBBVA 2018-2021). M.M.A. is funded by MICINN and the Spanish Government (no. PGC2018-094091-B-I00). D.C. was funded by Instituto de Salud Carlos III (no. JR1800003). S.M. is funded by PERIS (no. SLT006/17/00040). The wotk of R.R.G. is funded by MICINN, the Spanish Government (no. PID2019-104948RB-I00) and the BBVA Foundation. S.B and J.P. acknowledge support from the Spanish Ministry of Science, the EMBL partnership and the CESO and CERCA Program.

Introduction: Crizotinib is a first-in-class ALK tyrosine kinase inhibitor (TKI), which has proven its superiority over standard platinum-based chemotherapy for the first-line therapy of ALK-rearranged non-small cell lung cancer (NSCLC) patients. The development of acquired resistance to crizotinib represents an ongoing challenge with the central nervous system being one of the most common sites of relapse. Ceritinib and alectinib are approved second-generation ALK TKIs. Several novel ALK inhibitors, more potent and with different selectivity compared to crizotinib, are currently in development.Areas covered: This review will focus on new ALK inhibitors, currently in phase 1 or 2 clinical studies. We will also comment on the mechanisms of resistance to ALK inhibition and the strategies to delay or overcome resistance.Expert opinion: The therapeutic management of ALK-rearranged NSCLC has been greatly improved. Next-generation ALK inhibitors have shown differential potency against ALK rearrangements and ALK resistance mutations. The molecular profile of the tumor at the time of disease progression to crizotinib is crucial for the sequencing of novel ALK TKIs. Ongoing clinical studies will address key issues, including the optimal therapeutic algorithm and whether combinational approaches are more effective than single ALK inhibition for the outcome of ALK-rearranged NSCLC patients.

Alguns dels objectius de desenvolupament sostenible que proposa l’Agenda 2030 són: posar fi a la fam, assolir la seguretat alimentària i la millora de la nutrició i promoure l’agricultura sostenible. Assumir aquests reptes requereix l’aplicació dels avenços científics i tecnològics més punters per transformar un sector estratègic per a la nostra societat. Cal saber aplicar aquelles tecnologies que s’han desenvolupat en altres sectors per millorar la productivitat o, simplement, per introduir, a partir de la innovació, noves maneres de fer que alhora alleugerin les tasques manuals més feixugues. Ens apropem a un nou model emergent de gestió agrícola anomenat Smart Farm que es basa en un ús intensiu de les TIC i de sistemes automàtics i que, per tant, permet augmentar la qualitat i quantitat de producció i reduir l’esforç humà aplicat. En aquest article, ens centrarem en els aspectes més tecnològics que, aplicats al sector agroalimentari, poden generar uns canvis més disruptius.

BRAF V600 mutations have been found in 1&ndash, 2% of non-small-cell lung cancer (NSCLC) patients, with Food and Drug Administration (FDA) approved treatment of dabrafenib plus trametinib and progression free survival (PFS) of 10.9 months. However, 50&ndash, 80% of BRAF mutations in lung cancer are non-V600, and can be class II, with intermediate to high kinase activity and RAS independence, or class III, with impaired kinase activity, upstream signaling dependence, and consequently, sensitivity to receptor tyrosine kinase (RTK) inhibitors. Plasma cell-free DNA (cfDNA) of 185 newly diagnosed advanced lung adenocarcinoma patients (Spanish Lung Liquid versus Invasive Biopsy Program, SLLIP, NCT03248089) was examined for BRAF and other alterations with a targeted cfDNA next-generation sequencing (NGS) assay (Guardant360&reg, Guardant Health Inc., CA, USA), and results were correlated with patient outcome. Cell viability with single or combined RAF, MEK, and SHP2 inhibitors was assessed in cell lines with BRAF class I, II, and III mutations. Out of 185 patients, 22 had BRAF alterations (12%) of which seven patients harbored amplifications (32%) and 17 had BRAF mutations (77%). Of the BRAF mutations, four out of 22 (18%) were V600E and 18/22 (82%) were non-V600. In vitro results confirmed sensitivity of class III and resistance of class I and II BRAF mutations, and BRAF wild type cells to SHP2 inhibition. Concomitant MEK or RAF and SHP2 inhibition showed synergistic effects, especially in the class III BRAF-mutant cell line. Our study indicates that the class of the BRAF mutation may have clinical implications and therefore should be defined in the clinical practice and used to guide therapeutic decisions.

Introduction: Urine was shown to be a liquid biopsy source with high sensitivity for cfDNA mutation detection in both early- and late-stage cancer patients, outperforming plasma cfDNA and tissue samples in some patients. In addition, obtaining a urine sample is completely noninvasive. Limitations of using urine samples in the clinical setting include their unknown nucleic acid stability and the large total volume of one deposit, and therefore the presence of highly diluted cfDNA. Urine volume concentration, cfDNA stability, cfDNA quantity, and detection of wild-type (wt) EGFR alleles were explored to determine the future suitability of urine cfDNA as a diagnostic or prognostic biosource. Methods: First-void urine samples, with a higher fraction of nucleic acids due to overnight concentration, were collected from 5 healthy donors and 5 cancer patients. Cell-free urine samples were concentrated using Amicon filter tubes to determine differences in cfDNA concentration. In addition, we tested the ability to detect wild-type EGFR exon 19 in cfDNA from concentrated and unconcentrated urine samples using a peptide nucleic acid (PNA) Taqman Assay. Hereafter, stability of urine cfDNA was explored using variable storage temperatures and times and addition of a urine cfDNA preservative directly after sample collection. cfDNA was extracted using the automated QIAsymphony system with the DSP virus/pathogen kit and quantified using the Qubit dsDNA HS Assay in all experiments. Results: Amicon filter-based concentration of urine samples from 15 mL to 1.2 mL yielded a two-fold increase in cfDNA concentration. Importantly, while wild-type EGFR could not be detected in first-void urine using a PNA Taqman Assay, detection was possible using concentrated urine samples. Therefore, in all following experiments, samples were concentrated unless specified otherwise. Stability tests indicated that storage temperature matters when samples are not processed within two hours, with nearly doubled cfDNA concentrations found in samples stored at four degrees for 26 hours, compared to room temperature (RT). Addition of a cfDNA preservative yielded a six-fold higher cfDNA concentration after 26 hours of storage at RT, compared to samples without a preservative added. For this reason, we tested the use of only cfDNA preservative without concentrating the sample and found a four-fold increase in cfDNA concentration when adding only preservative, versus a 15-fold increase when using both after sample storage at RT for 24 hours. Conclusion: Patient urine samples should be either concentrated and processed within two hours, or a urine cfDNA preservative should be added directly after sample collection to prevent cfDNA degradation. Depending on the downstream application, samples where preservative was added may be concentrated to enhance the cfDNA yield, for example, when downstream NGS analysis will be performed. Citation Format: Jillian W. P. Bracht, Niki Karachaliou, Jordi Berenguer, Manuel Fernandez-Bruno, Mónica Garzón, Ana Gimenez-Capitan, Clara Mayo-de-las-Casas, Miguel Angel Molina-Vila, Rafael Rosell. Urine cell-free DNA (cfDNA) concentration and stability test for future clinical use [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B03.

Monotherapy with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) still leads to incomplete responses in most EGFR-mutation positive non-small cell lung cancer (NSCLC) patients, often due to acquired resistance through activation of parallel compensatory pathways. We have previously shown that co-targeting EGFR, signal transducer and activator of transcription 3 (STAT3), and Src-yes-associated protein 1 (YAP1) was highly synergistic in vitro and in vivo. In the present study, we treated EGFR-mutation positive cell lines with the combination of osimertinib plus a natural compound, pterostilbene, which has been reported to abrogate Src and STAT3 activation. Methods: Cell viability assays and immunoblotting were performed to reveal the mechanisms of action of pterostilbene, osimertinib and pterostilbene plus osimertinib in five EGFR-mutation positive NSCLC and one triple negative breast cancer (TNBC) cell lines. Results: Osimertinib plus pterostilbene yielded synergistic effects in all EGFR-mutation positive NSCLC cell lines investigated. Surprisingly, pterostilbene alone did not inhibit, nor downregulate Src phosphorylation in the EGFR-mutation positive NSCLC cell lines or the TNBC cell line, MDA-MB-231. However, the double combination of osimertinib plus pterostilbene reversed the osimertinib-induced STAT3, YAP1, and CUB domain-containing protein-1 (CDCP1) phosphorylation and slightly suppressed Src phosphorylation in PC9 and H1975 cells. Conclusion: The results of this study indicate that pterostilbene may be used to abrogate the activated resistance pathways of single osimertinib treatment in EGFR-mutation positive NSCLC. Future studies should focus on in vivo translation and confirmation of these results.

Background: The activation of multiple signaling pathways jeopardizes the clinical efficacy of EGFR tyrosine kinase inhibitors (TKIs) in EGFR-mutation positive non-small cell lung cancer (NSCLC). Integrin-1 inked kinase (ILK) regulates the interactions between tumor cells and extracellular environment to activate signaling pathways and promote cell proliferation, migration, and epithelial-mesenchymal transition. Src homology 2 domain-containing phosphatase 2 (SHP2) is essential for receptor tyrosine kinase signaling and mitogenactivated protein kinase (MAPK) pathway activation. Methods: We analyzed tumor ILK,13-receptor subunit glycoprotein 130 (gp130), SHP2, and stromal hepatocyte growth factor (HGF) and interleukin-6 (IL-6) mRNA expression in baseline tumor specimens of advanced EGFR-mutation positive NSCLC patients treated with EGFR TKIs. Results: ILK, when highly expressed, was an independent poor prognostic factor for the progression-free survival of the patients, both in the univariate (hazard ratio [HR for disease progression, 2.49; 95% Cl, 1.37-452; P .0020]) and in the multivariate (HR 3.74; 95% Cl, 1.33-10.56; P.0126) Cox regression model. Patients with high SHP2 expression had an almost 13-month shorter progression-free survival (P.0094) and an 18 month shorter overall survival (P.0182) in comparison to those with low SHP2 mRNA expression. Interpretation: The levels of ILK and SHP2 could be predictive for upfront combinatory therapy of EGFR TKIs plus SHP2 or ILK inhibitors. Fund: A grant from La Caixa Foundation, an Institute de Salud Carlos III grant (RESPONSE, PIE16/00011), an Institute de Salud Carlos III grant (PI14/01678), a Marie Sklodowska-Curie Innovative Training Networks European Grant (ELBA No 765492) and a Spanish Association Against Cancer (AECC) grant (PROYE18012ROSE). (C) 2018 The Authors. Published by Elsevier B.V.

Background: Osimertinib improve therapy for non-small cell lung cancer (NSCLC). However, invariable acquired resistance appears. Methods: MTT assay was used to analyze cell viability. Protein expression and activation was detected by Western blotting. In addition, the effects of heat shock protein 90 (Hsp90) inhibitors and osimertinib were studied in colony formation assays. Results: Our laboratory generated osimertinib resistant cell lines from PC9 cell line and overexpression or activation of several proteins was detected. Hsp90 inhibitors, ganetespib and luminespib, inhibited cell viability and colony formation in H1975, PC9 and PC9-derived osimertinib-resistant cell lines and combination of these inhibitors with osimertinib achieved to enhance this cell viability and colony formation inhibition. Luminespib downregulated the expression of the several proteins involved in osimertinib-resistance and the combination of this compound plus osimertinib caused an important decrease of expression of several of these proteins, such as Stat3, Yap, Akt, EGFR and Met. Osimertinib activated the phosphorylation of several membrane receptors and downstream molecules that was partially inhibited by luminespib. In addition, a lung cancer patient with an EGFR eon 20 mutation had a partial radiographic response to ganetespib. Conclusions: Hsp90 inhibitors and osimertinib exhibits a good efficiency to inhibit cell viability, colony formation and inhibits expression and activation of proteins involved in osimertinib-resistance and may represent an effective strategy for NSCLC with intrinsic resistance to osimertinib inhibition.

Epidermal growth factor receptor (EGFR)-mutation-positive non-small cell lung cancer (NSCLC) is incurable, despite high rates of response to EGFR tyrosine kinase inhibitors (TKIs). We investigated receptor tyrosine kinases (RTKs), Src family kinases and focal adhesion kinase (FAK) as genetic modifiers of innate resistance in EGFR-mutation-positive NSCLC. We performed gene expression analysis in two cohorts (Cohort 1 and Cohort 2) of EGFR-mutation-positive NSCLC patients treated with EGFR TKI. We evaluated the efficacy of gefitinib or osimertinib with the Src/FAK/Janus kinase 2 (JAK2) inhibitor, TPX0005 in vitro and in vivo. In Cohort 1, CUB domain-containing protein-1 (CDCP1) was an independent negative prognostic factor for progression-free survival (hazard ratio of 1.79, p = 0.0407) and overall survival (hazard ratio of 2.23, p = 0.0192). A two-gene model based on AXL and CDCP1 expression was strongly associated with the clinical outcome to EGFR TKIs, in both cohorts of patients. Our preclinical experiments revealed that several RTKs and non-RTKs, were up-regulated at baseline or after treatment with gefitinib or osimertinib. TPX-0005 plus EGFR TKI suppressed expression and activation of RTKs and downstream signaling intermediates. Co-expression of CDCP1 and AXL is often observed in EGFR-mutation-positive tumors, limiting the efficacy of EGFR TKIs. Co-treatment with EGFR TKI and TPX-0005 warrants testing., Graphical Abstract Unlabelled Image, Highlights • AXL and CDCP1 are co-expressed in treatment-naïve EGFR-mutation-positive NSCLC patients. • AXL and CDCP1 are related to shorter progression-free survival with EGFR inhibitors and shorter overall survival. • Src family kinases and YAP1 are regulatory nodes for AXL and CDCP1 expression. • The combination of EGFR TKI with TPX-0005 is synergistic in EGFR-mutation-positive lung tumors in culture and in vivo. We explore the molecular changes that occur after the application of an EGFR inhibitor in EGFR-mutation positive tumors. The tumors do not acquire secondary drivers to overcome a primary driver but, counter-regulatory nodes observable before treatment, are immediately made apparent by pathway-specific intervention. The expression of the receptor tyrosine kinase AXL and the transmembrane protein CDCP1 in baseline samples of EGFR-mutation positive NSCLC patients can provide us with information on the treatment outcome. The upfront combination of an EGFR inhibitor with a multikinase inhibitor, that controls the regulatory nodes for RTKs activation, is a therapeutic approach that deserves to be further explored.

Liquid biopsies have potential as tools for diagnosis, prognosis, and prediction of response to therapy. Herein, we will extensively review four liquid biosources, tumor-educated platelets (TEPs), cell-free DNA (cfDNA), circulating tumor cells (CTCs), and extracellular vesicles (EVs) and we will clarify their optimal application in non-small cell lung cancer (NSCLC) diagnosis and therapy.Liquid biopsies are a minimally invasive alternative to tissue biopsies-especially important in NSCLC patients-since tumor tissue is often unavailable or insufficient for complete genetic analysis. The main advantages of liquid biopsies include the possibility for repeated sampling, the lower cost, and the fact that they can reflect the complete molecular status of the patient better than a single-site biopsy. This is specifically important for lung adenocarcinoma patients since the detection of specific genetic alterations can predict response to targeted therapies. Molecular analysis is currently cardinal for therapy decision-making and disease monitoring in lung cancer patients. Liquid biopsies can make easier our daily clinical practice and if prospectively tested and validated may serve as a means for lung cancer early detection.

Background MET addicted tumors are known to show very short response to single MET inhibition. In MET addicted cells, MET inhibition leads to activation of proviral integration site for Moloney murine leukemia virus-1 (PIM1). PIM1 and SRC can regulate the expression of receptor tyrosine kinases (RTKs), potentially being responsible of resistance to MET inhibition. We previously showed that the dual inhibition of MET and PIM1 with class I MET inhibitors and the pan-PIM inhibitor AZD1208, as well as MET and SRC (with dasatinib), is synergistic in MET addicted cell lines. Methods We evaluated the activity of class I and class II MET inhibitors, MET/SRC/CSF1R inhibitor TPX-02226 and pan-PIM inhibitors AZD1208 and PIM447 in four MET addicted cell lines: 2 MET amplified lung cancer cell lines (EBC1 and H1993), 1 MET exon 14 mutant cell line (Hs746T) and 1 MET exon 7-8 splicing variant cell line (E98). We assessed the effect of the dual inhibition of MET and PIM, and the triple inhibition of MET, SRC and PIM in cell viability by combining the TPX-02226 with the pan-PIM inhibitors. Results All cell lines were sensitive to class I-II MET inhibitors (IC50s in nM range) except for cabozantinib in H1993. Only the MET amplified cell lines were sensitive to TPX-02226. All cell lines were resistant to PIM inhibition. The combination of class I MET inhibitors with PIM447 showed stronger synergism in the MET amplified cell lines, compared to the combination with AZD1208. Class I-II MET inhibitors and PIM inhibitor combination showed only additive effect in exon 7-8 cell line E98, while the combination of TPX-02226 and PIM447 was strongly synergistic. Conclusions We identified potential role of PIM inhibition in MET amplified tumors. Even though single agent TPX-02226, AZD1208 or PIM447 showed no activity and dual MET/PIM inhibition showed only additive effect, the triple inhibition of MET, SRC and PIM was strongly effective in MET exon 7-8 splicing variant, suggesting a crucial role of SRC-PIM interaction in this still not well recognized MET addicted tumor. Further investigation on this triple inhibition is ongoing in MET addicted cell lines and role of co-presence of MET and PIM1 and/or SRC alterations in tumor samples. Legal entity responsible for the study IGTP, Germans Trias i Pujol Research Institute, Badalona, Barcelona, Spain. Funding Fundacio Obra Social “La Caixa”. Disclosure J.J. Cui: Leadership role, Officer / Board of Directors: TP Therapeutics. N. Karachaliou: Officer / Board of Directors: Merck KgaA. All other authors have declared no conflicts of interest.

The therapeutic possibilities for patients with metastatic melanoma have changed due to the development of targeted therapies that inhibit oncogenic signaling pathways as well as immune modulating therapies that unleash the patient antitumor immunity. These therapeutic changes have impressively increased the median overall survival of the patients. Considering the dramatic but transient responses that occur with targeted therapies for a subgroup of patients and the durable responses that can be achieved with immunotherapy in a subset of patients, a lot of effort is ongoing for the clinical development of combinations of these two therapeutic approaches. Herein we discuss the existing preclinical and clinical data for the combination of targeted therapies and immunotherapy focusing mainly on melanoma and non-small cell lung cancer (NSCLC).

e24001Background: p21-activated kinase 1 (PAK1) stimulates growth and metastasis in several types of tumors, including NSCLC. Protein kinase C iota (PKCi) is highly expressed in NSCLC and regulates...

e24003Background: Several non receptor tyrosine kinases (RTKs) activated by EGFR tyrosine kinase inhibitors (TKI) have been identified. Src family kinases (SFKs), FAK and STAT3 protein and mRNA lev...

Nmyc is a potent regulator of cell cycle in cerebellar granular neuron precursors (CGNPs) and has been proposed to be the main effector of Shh (Sonic hedgehog) proliferative activity. Nmyc ectopic expression is sufficient to promote cell autonomous proliferation and can lead to tumorigenesis. Bone morphogenetic protein 2 (BMP2) antagonizes Shh proliferative effect by promoting cell cycle exit and differentiation in CGNPs. Here we report that BMP2 opposes Shh mitogenic activity by blocking Nmyc expression. We have identified TIEG-1 (KLF10) as the intermediary factor that blocks Nmyc expression through the occupancy of the Sp1 sites present in its promoter. We also demonstrate that TIEG-1 ectopic expression in CGNPs induces cell cycle arrest that can lead to apoptosis but fails to promote differentiation. Moreover, TIEG-1 synergizes with BMP2 activity to terminally differentiate CGNPs and independent differentiator signals such as dibutyryl cAMP and prevents apoptosis in TIEG-1 arrested cells. All together, these data strongly suggest that the BMP2 pathway triggers cell cycle exit and differentiation as two separated but coordinated processes, where TIEG-1 acts as a mediator of the cell cycle arrest. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc., This work was supported by Ministerio de Educación y Ciencia Grant BFU2005-01599.

// R. Jonas A. Nilsson 1,2,3,* , Niki Karachaliou 4,* , Jordi Berenguer 1 , Ana Gimenez-Capitan 5 , Pepijn Schellen 1,3 , Cristina Teixido 5 , Jihane Tannous 6 , Justine L. Kuiper 7 , Esther Drees 1 , Magda Grabowska 1 , Marte van Keulen 6 , Danielle A. M. Heideman 8 , Erik Thunnissen 8 , Anne-Marie C. Dingemans 9 , Santiago Viteri 4 , Bakhos A. Tannous 6 , Ana Drozdowskyj 10 , Rafael Rosell 4,5,11,12,** , Egbert F. Smit 7,** and Thomas Wurdinger 1,3,6,** 1 Cancer Center Amsterdam, Department of Neurosurgery, VU University Medical Center, Amsterdam, The Netherlands 2 Department of Radiation Sciences, Oncology, Umea University, Umea, Sweden 3 ThromboDx B.V., Amsterdam, The Netherlands 4 Translational Research Unit, Dr, Rosell Oncology Institute, Quiron Dexeus University Hospital, Barcelona, Spain 5 Pangaea Biotech SL, Barcelona, Spain 6 Department of Neurology, Massachusetts General Hospital and Neuroscience Program, Harvard Medical School, Boston, MA, USA 7 Cancer Center Amsterdam, Department of Pulmonary Diseases, VU University Medical Center, Amsterdam, The Netherlands 8 Cancer Center Amsterdam, Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands 9 Department of Pulmonary Diseases, Maastricht University Medical Center, Maastricht, The Netherlands 10 Pivotal, Madrid, Spain 11 Catalan Institute of Oncology, Hospital Germans Trias i Pujol, Barcelona, Spain 12 Molecular Oncology Research (MORe) Foundation, Barcelona, Spain * These two authors are co-first authors of the manuscript ** These three authors are co-senior authors of the manuscript Correspondence to: Thomas Wurdinger, email: // Keywords : diagnostics, NSCLC, liquid biopsies, platelets, EML4-ALK Received : August 23, 2015 Accepted : October 06, 2015 Published : November 02, 2015 Abstract Purpose: Non-small-cell lung cancers harboring EML4-ALK rearrangements are sensitive to crizotinib. However, despite initial response, most patients will eventually relapse, and monitoring EML4-ALK rearrangements over the course of treatment may help identify these patients. However, challenges associated with serial tumor biopsies have highlighted the need for blood-based assays for the monitoring of biomarkers. Platelets can sequester RNA released by tumor cells and are thus an attractive source for the non-invasive assessment of biomarkers. Methods: EML4-ALK rearrangements were analyzed by RT-PCR in platelets and plasma isolated from blood obtained from 77 patients with non-small-cell lung cancer, 38 of whom had EML4-ALK-rearranged tumors. In a subset of 29 patients with EML4-ALK-rearranged tumors who were treated with crizotinib, EML4-ALK rearrangements in platelets were correlated with progression-free and overall survival. Results: RT-PCR demonstrated 65% sensitivity and 100% specificity for the detection of EML4-ALK rearrangements in platelets. In the subset of 29 patients treated with crizotinib, progression-free survival was 3.7 months for patients with EML4-ALK+ platelets and 16 months for those with EML4-ALK− platelets (hazard ratio, 3.5; P = 0.02). Monitoring of EML4-ALK rearrangements in the platelets of one patient over a period of 30 months revealed crizotinib resistance two months prior to radiographic disease progression. Conclusions: Platelets are a valuable source for the non-invasive detection of EML4-ALK rearrangements and may prove useful for predicting and monitoring outcome to crizotinib, thereby improving clinical decisions based on radiographic imaging alone.

El 3 d’abril de 2010 es va produir la fi de les emissions de televisió analògica a l’Estat espanyol. És el que es coneix com l’apagada analògica. De llavors ençà s’inicià una nova etapa en la radiodifusió de la televisió fent ús d’un nou sistema de senyals i modulacions digitals que és el que anomenem Televisió Digital Terrestre o TDT. Es posava fi a una etapa d’emissions regulars de televisió analògica que a l’estat havien començat el 1956, tot i que les primeres proves es van fer a Barcelona a la Fira de Mostres del 1948. Una etapa que sempre es va caracteritzar per garantir la compatibilitat amb els receptors existents del senyal de televisió, quan aquest anava incorporant al llarg dels anys, millores tecnològiques com ara el color o el so digital.

During cerebellum development, Sonic hedgehog (Shh)-induced proliferation of cerebellar granular neuronal precursors (CGNPs) is potently inhibited by bone morphogenetic proteins (BMPs). We have previously reported the upregulation of TIEG-1 and Mash1, two antimitotic factors that modulate MYCN transcription and N-Myc activity, in response to BMP2. To gain further insight into the BMP antimitotic mechanism, we used microRNA (miRNA) arrays to compare the miRNAs of CGNPs proliferating in response to Shh with those of CGNPs treated with Shh plus BMP2. The array analysis revealed that miRNA 11 (miR-22) levels significantly increased in cells treated with BMP2. Additionally, in P7 mouse cerebellum, miR-22 distribution mostly recapitulated the combination of BMP2 and BMP4 expression patterns. Accordingly, in CGNP cultures, miR-22 overexpression significantly reduced cell proliferation, whereas miR-22 suppression diminished BMP2 antiproliferative activity. In contrast to BMP2, miR-22 did not induce neural differentiation but instead significantly increased cell cycle length. Consistent with the central role played by N-myc on CGNP proliferation, Max was revealed as a direct target of miR-22, and miR-22 expression caused a significant reduction of Max protein levels and N-myc/Max-dependent promoter activity. Therefore, we conclude that, in addition to the previously described mechanisms, miR-22 plays a specific role on downstream BMPs through cerebellum growth, Work in S.P.’s laboratory is supported by the Spanish Ministry of Education grants BFU2008-02424/BFI and BFU2011-24099.

Cinnamyl alcohol dehydrogenase (CAD) is a key enzyme involved in the last step of monolignol biosynthesis. The effect of CAD down-regulation on lignin production was investigated through a transgenic approach in maize. Transgenic CAD-RNAi plants show a different degree of enzymatic reduction depending on the analyzed tissue and show alterations in cell wall composition. Cell walls of CAD-RNAi stems contain a lignin polymer with a slight reduction in the S-to-G ratio without affecting the total lignin content. In addition, these cell walls accumulate higher levels of cellulose and arabinoxylans. In contrast, cell walls of CAD-RNAi midribs present a reduction in the total lignin content and of cell wall polysaccharides. In vitro degradability assays showed that, although to a different extent, the changes induced by the repression of CAD activity produced midribs and stems more degradable than wild-type plants. CAD-RNAi plants grown in the field presented a wild-type phenotype and produced higher amounts of dry biomass. Cellulosic bioethanol assays revealed that CAD-RNAi biomass produced higher levels of ethanol compared to wild-type, making CAD a good target to improve both the nutritional and energetic values of maize lignocellulosic biomass., This work was supported by the Spanish ‘Ministerio de Ciencia e Innovación’ (AGL2008–05157), the European Community (‘COPOL’ project: integrated control of polysaccharide and lignin biosynthesis to improve cellulose content and availability and fibre quality, QLK5-CT-2000–01493), and the CONSOLIDER-INGENIO program (CSD2007–00036). S.F. was financed by an I3P contract from the ‘Consejo Superior de Investigaciones Científicas’. D.C.-R. was initially financed by the Spanish ‘Ministerio de Educación y Ciencia’ (‘Ramón y Cajal’ Program). A.E. was financed by DGICYT (CGL2008–02470BOS) and Junta de Castilla y León (LE004A10-2). This work was carried out within the framework of the ‘Xarxa de Referència de Biotecnologia’ (XarBa) from the Autonomous Government of Catalonia.

Cerebellar granular neuronal precursors (CGNPs) proliferate in response to the mitogenic activity of Sonic hedgehog (Shh), and this proliferation is negatively regulated by activation of cAMP-dependent protein kinase (PKA). In the basal state, the PKA catalytic subunits (C-PKA) are inactive because of their association with the regulatory subunits (R-PKA). As the level of cAMP increases, it binds to R-PKA, displacing and thereby activating the C-PKA. Here we report that, in the presence of Shh, inactive C-PKA accumulates at the cilium base of proliferative CGNPs whereas removal of Shh triggers the activation of PKA at this particular location. Furthermore, we demonstrate that the anchoring of the PKA holoenzyme to the cilium base is mediated by the specific binding of the type II PKA regulatory subunit (RII-PKA) to the A-kinase anchoring proteins (AKAPs). Disruption of the interaction between RII-PKA and AKAPs inhibits Shh activity and, therefore, blocks proliferation of CGNP cultures. Collectively, these results demonstrate that the pool of PKA localized to the cilium base of CGNP plays an essential role in the integration of Shh signal transduction., Work in S. P.'s laboratory is supported by the Spanish Ministry of Education Grant BFU200802424/BFI.

El pdf del artículo es el manuscrito de autor., Methylmercury (MeHg) is an environmental neurotoxicant whose molecular mechanisms underlying toxicity remain elusive. Here we investigated molecular events involved in MeHg-induced neurotoxicity in cultured cerebellar granule cells (CGCs) as well as potential protective strategies for such toxicity. Glutathione peroxidase (GPx-1) activity was significantly (p = 0.0017) decreased at 24 h before MeHg-induced neuronal death (day in vitro 4). This event was related to enhanced susceptibilities to hydrogen- or tert-butyl-peroxides and increased lipid peroxidation. However, intracellular calcium levels, glutamate uptake and glutathione levels, as well as glutathione reductase and catalase activities, were not changed by MeHg exposure at this time-point. Probucol (PB), a lipid-lowering drug, displayed a long-lasting protective effect against MeHg-induced neurotoxicity. The beneficial effects of PB were correlated to increased GPx-1 activity and decreased lipid peroxidation. The protection afforded by PB was significantly higher when compared to the antioxidants ascorbic acid and trolox. In vitro studies with the purified GPx-1 proved that MeHg inhibits and PB activates the enzyme activity. Overexpression of GPx-1 prevented MeHg-induced neuronal death. These data indicate that (i) GPx-1 is an important molecular target involved in MeHg-induced neurotoxicity and (ii) PB, which increases GPx-1 activity in CGCs, induces enduring protection against such toxicity. The results bring out new insights on the potential therapeutic strategies for poisonings to MeHg and other pathological conditions related to increased production and/or decreased detoxification of peroxides., This study was supported by the Grant FIS PI061212 and 2005-SGR-00826 (Ministry of Health and Generalitat de Catalunya, respectively, Spain). Marcelo Farina was recipient of a post- doctoral fellowship (Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq/201362/2007-4) and received financial support from CNPq (479239/2007-0) and FAPESC (Jovens Pesquisadores - FAPESC/CNPq 04/2007).

Et presentem el desè monogràfic que hem preparat des de l’Associació i el Col·legi d’Enginyers Industrials de Catalunya. En aquesta ocasió hem volgut posar el focus d’atenció en un sector molt important per al desenvolupament de la nostra societat: l’aeronàutica. Es tracta d’un sector amb un futur prometedor si som capaços de vehicular tots els esforços que són necessaris per donar-li impuls. De fet, el sector aeronàutic va ser molt important a Catalunya cap al 1930; amb posterioritat, determinades voluntats polítiques en van frenar la continuïtat. Aquest monogràfic coincideix amb un any important per al nostre país: la inauguració de la nova T1 del Prat, que sens dubte esdevé un nou impuls per al nostre territori. Amb la intenció de fer una radiografia d’aquest sector, presentem un recull d’articles on s’aglutinen una gran quantitat d’idees exposades per persones enteses en la matèria, i una diagnosi redactada amb gran rigor per un company que fa molts anys que treballa en el sector. El nostre objectiu és contribuir a generar debat social entre els ciutadans, empreses, institucions i la resta d’agents implicats. Esperem que us sembli tan interessant com a nosaltres i aprofitem aquestes línies per agrair la participació de tots els que han fet possible aquesta publicació.

Background: Novel targeted therapies have been successfully used against subgroups of non-small-cell lung cancer (NSCLC) patients, however, despite initial good response the cancer will eventually relapse. One of those subgroups harbors a rearranged EML4-ALK fusion gene that makes them responsive to crizotinib treatment, but therapy resistance often soon occurs. Real-time monitoring of rearrangement status over the course of treatment will help identify patients showing therapy resistance, but monitoring through serial tumor biopsies has been an obstacle. Therefore, new blood-based ´liquid biopsy´ platforms need to be developed to monitoring biomarkers in the circulation. Here we present one platform using the ability of platelets to sequester RNA released by tumor cells and represent an attractive source for non-invasive biomarker assessment. Methods: EML4-ALK rearrangements were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) in platelets and plasma isolated from blood obtained from 77 NSCLC patients, 38 of whom had EML4-ALK-rearranged tumors. In a subset patients (n = 29) that were treated with crizotinib, progression-free survival (PFS) and overall survival (OS) were correlated with presence of EML4-ALK rearrangements in platelets. Results: The study was designed with three parallel objectives: firstly to determine the sensitivity and specificity of detecting EML4-ALK rearrangements in platelets; secondly, to examine the potential impact of EML4-ALK rearrangement in platelets on outcome to crizotinib; thirdly, to test the feasibility of monitoring patients throughout treatment with EML4-ALK rearrangement assessment in platelets. Detection of EML4-ALK rearrangements in platelets demonstrated 65% sensitivity and 100% specificity. The PFS in patients treated with crizotinib was 3.7 months in patients with a positive EML4-ALK platelet status compared to 16 months for a negative EML4-ALK status (hazard ratio, 3.5; P = 0.02). Furthermore, longitudinal monitoring of EML4-ALK rearrangements in platelets was feasible, as demonstrated in an index patient where crizotinib resistance was observed two months prior to radiographic disease progression. Conclusions: Platelets may provide a useful source for non-invasive assessment of EML4-ALK rearrangements and may prove useful for predicting outcome to crizotinib. Serial analyses of EML4-ALK rearrangements in platelets may help improve clinical decisions based on radiographic imaging alone by detecting resistance to therapy sooner. Citation Format: Jonas A. Nilsson, Niki Karachaliou, Pepijn Schellen, Ana Gimenez-Capitan, Jordi Berenguer, Cristina Teixido, Justine L. Kuiper, Esther Drees, Magda Grabowska, Marte van Keulen, Jihane M. Tannous, Danielle A.M. Heideman, Erik Thunnissen, Anne-Marie C. Dingemans, Santiago Viteri, Bakhos A. Tannous, Ana Drozdowskyj, Rafael Rosell, Egbert F. Smit, Thomas Wurdinger. Monitoring rearrangement of EML4-ALK in blood platelets predicts outcome to crizotinib treatment in non-small-cell lung cancer patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-053. doi:10.1158/1538-7445.AM2015-LB-053

8082 Background: Non-small-cell lung cancer (NSCLC) with EML4-ALK rearrangements is sensitive to crizotinib. However, despite initial response most patients (p) will eventually relapse and monitori...

El llibre presenta el desenvolupament d'un conjunt d'experiències de laboratori destinades a posar en evidència els aspectes més característics de la tecnologia de la radiofreqüència aplicada a la telecomunicació, tant pel que fa al disseny de circuits i la caracterització de subsistemes com a l'ús de la instrumentació que li és pròpia, complementats amb una anàlisi i un desenvolupament teòrics. I inclou també un primer capítol que ofereix una visió global dels diferents serveis i sistemes de telecomunicació, com també un altre dedicat a mesures d'EMC-EMI radiades. Finalment, s'hi adjunta un disquet que conté la documentació de fabricació dels circuits impresos proposats, més uns programes d'anàlisi i síntesi de línies de transmissió microstrip.

Summary Most cancer cells release heterogeneous populations of extracellular vesicles (EVs) containing proteins, lipids, and nucleic acids. In vitro experiments showed that EV uptake can lead to transfer of functional mRNA and altered cellular behavior. However, similar in vivo experiments remain challenging because cells that take up EVs cannot be discriminated from non-EV-receiving cells. Here, we used the Cre-LoxP system to directly identify tumor cells that take up EVs in vivo. We show that EVs released by malignant tumor cells are taken up by less malignant tumor cells located within the same and within distant tumors and that these EVs carry mRNAs involved in migration and metastasis. By intravital imaging, we show that the less malignant tumor cells that take up EVs display enhanced migratory behavior and metastatic capacity. We postulate that tumor cells locally and systemically share molecules carried by EVs in vivo and that this affects cellular behavior., Graphical Abstract, Highlights • Visualization of extracellular vesicle exchange between tumor cells in living mice • Direct in vivo evidence of local and systemic mRNA exchange between tumor cells • The exchange of biomolecules is mediated through extracellular vesicles • Metastatic behavior can be phenocopied through extracellular vesicle exchange, Intravital imaging experiments reveal the extracellular vesicle-mediated exchange of molecules important for metastasis between tumor cells in living mice. This exchange results in metastatic properties being conferred to the receiving cell.

Este segundo volumen de la historia de Telefónica arroja una mirada de gran angular sobre el desarrollo de la compañía desde los años 70 hasta los primeros años del siglo xxi, unas décadas marcadas por la apertura de mercados, la globalización y la expansión de las telecomunicaciones. En este contexto, la internacionalización de la compañía se convirtió en un ejemplo de estrategia internacional, con especial relevancia en los mercados de América Latina. Los hechos incluidos en esta obra son el resultado de un análisis exhaustivo, que incluye una amplia tarea de documentación y reflexión teórica, para la que se han consultado variadas fuentes documentales hasta ahora no exploradas y realizado entrevistas a los principales protagonistas de esta época. Para ello, hemos contado con la autoría de expertos actuales provenientes de distintas disciplinas académicas, que han sabido relacionar las variables tecnológicas, políticas, económicas y sociales que influyeron en el devenir de la Compañía. Todo ello ha contribuido a forjar una obra historiográfica de indudable recorrido que permite trazar el proceso de internacionalización de Telefónica desde un novedoso punto de vista.