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The humanized anti-p185(HER2) monoclonal antibody trastuzumab has been shown to effectively inhibit the growth of HER2-overexpressing breast cancer cells in vivo and in vitro. The treatment of cancer cells with trastuzumab results in downregulation of the HER2 receptor. Further downstream cellular events include the accumulation of the cyclin-dependent kinase inhibitor p27 and cell cycle arrest. In vivo, trastuzumab induces antibody-dependent cellular cytotoxicity. Trastuzumab also inhibits constitutive HER2 cleavage/shedding mediated by metalloproteases. The ability of trastuzumab to inhibit HER2 cleavage may correlate with the clinical anticancer activity of the multifunctional HER2-targeting antibody.

The role of each residue of the potato carboxypeptidase inhibitor (PCI) C-terminal tail, in the interaction with carboxypeptidase A (CPA), has been studied by the analysis of two main kinds of site-directed mutants: the point substitution of each C-terminal residue by glycine and the sequential deletions of the C-terminal residues. The mutant PCI‐CPA interactions have been characterized by the measurement of their inhibition constant, Ki ,i n several cases, by their kinetic association and dissociation constants determined by presteady-state analysis, and by computational approaches. The role of Pro36 appears to be mainly the restriction of the mobility of the PCI C-tail. In addition, and unexpectedly, both Gly35 and Pro36 have been found to be important for folding of the protein core. Val38 has the greatest enthalpic contribution to the PCI‐CPA interaction. Although Tyr37 has a minor contribution to the binding energy of the whole inhibitor, it has been found to be essential for the interaction with the enzyme following the cleavage of the C-terminal Gly39 by CPA. The energetic contribution of the PCI secondary binding site has been evaluated to be about half of the total free energy of dissociation of the PCI‐CPA complex.

Epidermal growth factor (EGF) and its receptor (EGFR) are involved in many aspects of the development of carcinomas, including tumor cell growth, vascularization, invasiveness, and metastasis. Because EGFR has been found to be overexpressed in many tumors of epithelial origin, it is a potential target for antitumor therapy. Here we report that potato carboxypeptidase inhibitor (PCI), a 39-amino acid protease inhibitor with three disulfide bridges, is an antagonist of human EGF. It competed with EGF for binding to EGFR and inhibited EGFR activation and cell proliferation induced by this growth factor. PCI suppressed the growth of several human pancreatic adenocarcinoma cell lines, both in vitro and in nude mice. PCI has a special disulfide scaffold called a T-knot that is also present in several growth factors including EGF and transforming growth factor alpha. PCI shows structural similarities with these factors, a fact that can explain the antagonistic effect of the former. This is the first reported example of an antagonistic analogue of human EGF.

Purpose: Concomitant genetic alterations could account for transient clinical responses to tyrosine kinase inhibitors of the EGF receptor (EGFR) in patients harboring activating EGFR mutations. Experimental Design: We have evaluated the impact of pretreatment somatic EGFR T790M mutations, TP53 mutations, and Bcl-2 interacting mediator of cell death (BCL2L11, also known as BIM) mRNA expression in 95 patients with EGFR-mutant non–small-cell lung cancer (NSCLC) included in the EURTAC trial (trial registration: NCT00446225). Results: T790M mutations were detected in 65.26% of patients using our highly sensitive method based on laser microdissection and peptide-nucleic acid-clamping PCR, which can detect the mutation at an allelic dilution of 1 in 5,000. Progression-free survival (PFS) to erlotinib was 9.7 months for those with T790M mutations and 15.8 months for those without, whereas among patients receiving chemotherapy, it was 6 and 5.1 months, respectively (P < 0.0001). PFS to erlotinib was 12.9 months for those with high and 7.2 months for those with low/intermediate BCL2L11 expression levels, whereas among chemotherapy-treated patients, it was 5.8 and 5.5 months, respectively (P = 0.0003). Overall survival was 28.6 months for patients with high BCL2L11 expression and 22.1 months for those with low/intermediate BCL2L11 expression (P = 0.0364). Multivariate analyses showed that erlotinib was a marker of longer PFS (HR = 0.35; P = 0.0003), whereas high BCL2L11 expression was a marker of longer PFS (HR = 0.49; P = 0.0122) and overall survival (HR = 0.53; P = 0.0323). Conclusions: Low-level pretreatment T790M mutations can frequently be detected and can be used for customizing treatment with T790M-specific inhibitors. BCL2L11 mRNA expression is a biomarker of survival in EGFR-mutant NSCLC and can potentially be used for synthetic lethality therapies. Clin Cancer Res; 20(7); 2001–10. ©2014 AACR.

Somatic mutations in protein kinases (PKs) are frequent driver events in many human tumors, while germ-line mutations are associated with hereditary diseases. Here we present Kin-driver, the first database that compiles driver mutations in PKs with experimental evidence demonstrating their functional role. Kin-driver is a manual expert-curated database that pays special attention to activating mutations (AMs) and can serve as a validation set to develop new generation tools focused on the prediction of gain-of-function driver mutations. It also offers an easy and intuitive environment to facilitate the visualization and analysis of mutations in PKs. Because all mutations are mapped onto a multiple sequence alignment, analogue positions between kinases can be identified and tentative new mutations can be proposed for studying by transferring annotation. Finally, our database can also be of use to clinical and translational laboratories, helping them to identify uncommon AMs that can correlate with response to new antitumor drugs. The website was developed using PHP and JavaScript, which are supported by all major browsers; the database was built using MySQL server. Kin-driver is available at: http://kin-driver.leloir.org.ar/ Fil: Simonetti, Franco Lucio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina Fil: Tornador, Cristian. Universitat Pompeu Fabra; España Fil: Nabau Moretó, Nuria. Universitat de Barcelona. Facultat de Biologia; España Fil: Molina Vila, Miguel A.. Hospital Universitari Dexeus; España Fil: Marino Buslje, Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina

Objectives: Vorinostat or suberoylanilide hydroxamic acid (SAHA) is a novel histone deacetylase inhibitor with demonstrated antiproliferative effects due to drug-induced accumulation of acetylated proteins, including the heat shock protein 90. We prospectively studied the activity of vorinostat plus erlotinib in EGFR-mutated NSCLC patients with progression to tyrosine kinase inhibitors. Patients and methods: We conducted this prospective, non-randomized, multicenter, phase I/II trial to evaluate the maximum tolerated dose, toxicity profile and efficacy of erlotinib and vorinostat. Patients with advanced NSCLC harboring EGFR mutations and progressive disease after a minimum of 12 weeks on erlotinib were included. The maximum tolerated dose of vorinostat plus erlotinib was used as recommended dose for the phase II (RDP2) to assess the efficacy of the combination. The primary end point was progression-free-survival rate at 12 weeks (PFSR12w). Pre-treatment plasma samples were required to assess T790M resistant mutation. Results: A total of 33 patients were enrolled in the phase I-II trial. The maximum tolerated dose was erlotinib 150 mg p.o., QD, and 400 mg p.o., QD, on days 1-7 and 15-21 in a 28-day cycle. Among the 25 patients treated at the RDP2, the most common toxicities included anemia, fatigue and diarrhea. No responses were observed. PFSR12w was 28% (IC95%: 18.0-37.2); median progression-free survival (PFS) was 8 weeks (IC 95%: 7.43-8.45) and overall survival (OS) 10.3 months (95% CI: 2.4-18.1). Conclusion: Full dose of continuous erlotinib with vorinostat 400 mg p.o., QD on alternative weeks can be safely administered. Still, the combination has no meaningful activity in EGFR-mutated NSCLC patients after TKI progression. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Introduction Lung adenocarcinoma patients harboring EGFR activating mutations attain improved progression-free survival (PFS) with treatment with epidermal growth factor receptor tyrosine kinase inhibitors. However, patients ultimately relapse, indicating that other genetic factors could influence outcome in such patients. We hypothesized that PFS could be influenced by the expression of genes in DNA repair pathways. Methods We examined the mRNA expression of C terminus-binding protein–interacting protein and Lin11 , Isl-1 , and Mec-3 domain only 4 ( LMO4 ) in pretreatment tumor samples from 91 erlotinib-treated advanced non–small-cell lung cancer patients with EGFR mutations in whom breast cancer gene 1 ( BRCA1 ) expression and the concomitant presence of the EGFR T790M mutation had previously been assessed. Gene expression was analyzed by polymerase chain reaction, using β-actin as endogenous gene. Results were correlated with PFS and overall survival. Results In patients with low LMO4 levels, PFS was 13 months, whereas it was not reached for those with high LMO4 levels ( p = 0.03). In patients with low levels of both BRCA1 and LMO4 , PFS was 19 months whereas it was not reached in those with low BRCA1 and high LMO4 mRNA levels ( p = 0.04). In patients with high BRCA1 and low LMO4 levels, PFS was 8 months, whereas it was 18 months in those with high levels of both genes ( p = 0.03). Conclusions Low BRCA1 and high LMO4 levels were associated with longer PFS to erlotinib. Baseline assessment of BRCA1 and LMO4 mRNA expression can help predict outcome to erlotinib.

Introduction: Discoidin domain receptor 2 (DDR2) is a receptor tyrosine kinase (RTK) activated by collagen I and identified as an oncogenic driver in squamous cell carcinoma of the lung (SCC) (1). DDR2 mutations have been reported in 4% of SCC patients and at lower frequencies in lung adenocarcinoma, gastric, breast and brain tumors (2). In addition, DDR2 is expressed during epithelial to mesenchymal transition (EMT) (3) and acts as an upstream activator of SHP2. SHP2 in turn is a key signaling effector which leads to activation of multiple signaling pathways (4). DDR2 has recently emerged as a potential therapeutic target in DDR2-mutated tumors and drugs such as dasatinib are being tested in this setting. Results and discussion: PB1 is a small organic molecule with a simple synthesis pathway that showed strong inhibitory activity against wild type (wt) and mutated purified DDR2 protein. The compound was subsequently tested in a panel of tumor cell lines from different origins and histologies. PB1 showed good antiproliferative activity: IC50 Conclusion: PB1 shows significant antitumor activity in preclinical models of mutated and wt DDR2 in SCC of the lung and some KRAS mutated adenocarcinoma cell lines. PB1 is also active against tumor cells with secondary resistance to dasatinib. References: 1) Hammerman et al. Cancer Discovery. 2011 June; 1: 78-89. 2) Terai et al. ACS Chem Biol. 2015 Sep; Epub ahead of print. 3) Walsh et al. Matrix Biol. 2011 May; 30(4): 243-247. 4) Iwai et al. Biochem. J. (2013) 454, 501-513. Citation Format: Silvia Garcia-Roman, Miguel Angel Molina, José Ignacio Borrell, Raimon Puig de la Bellacasa, Daniela Morales, Jordi Bertran, Ana Gimenez, Niki Karachaliou, Rafael Rosell. PB1, a DDR2 inhibitor with antitumor activity in preclinical models of squamous cell carcinoma and KRAS-mutated adenocarcinoma of the lung. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4802.

Intrinsic or acquired resistance limits the clinical effectiveness of EGFR tyrosine kinase inhibitors (TKIs) for non-small cell lung cancer (NSCLC) patients (p) with EGFR mutations. One of the signaling mediators downstream of activated EGFR is signal transducer and activator of transcription 3 (STAT3). Not only does gefitinib not inhibit STAT3, but it also augments STAT3 tyrosine phosphorylation. EGFR blockade enriches lung cancer stem cells (CSCs) through NOTCH3-dependent signaling. A co-receptor of IL-6 (gp130) associates with Src and triggers activation of YAP and NOTCH. Our study is designed with three parallel objectives: firstly, to demonstrate that single EGFR TKI treatment cannot abrogate STAT3 and Src in EGFR mutant NSCLC cell lines; secondly, to examine whether the combination of gefitinib with compounds that target STAT3, (TPCA-1) and Src (saracatinib), suppresses the mechanisms of resistance; thirdly, to identify biomarkers in clinical tumor samples that may help us predict the outcome of EGFR TKIs and design effective combination therapies. Cell viability assay (MTT), western blotting, quantitative-real time PCR (qRT-PCR) and aldefluor assay-flow cytometry were used. We found that gefitinib increases pSTAT3 Y705 in PC-9 cells (that harbor the exon 19 deletion) in a time- and dose-dependent manner. Nine days after gefitinib treatment STAT3 mRNA level was significantly elevated. PC-9 cells showed dramatic increase in the fraction of ALDH+ cells upon treatment with gefitinib. TPCA-1 increased sensitivity to gefitinib in the PC-9 cells. Combination of gefitinib with TPCA-1 abrogated pSTAT3 Y705 but neither inhibited pPaxillin Y118 (Src induced) and pYAP S127 nor prevented the increment in the ALDH+ CSCs subpopulation. The triple combination of gefitinib, TPCA-1 and saracatinib was highly synergistic and abrogated pSTAT3 Y705, pPaxillin Y118 and pYAP S127. We performed qRT-PCR at baseline tumor samples of 64 EGFR mutant NSCLC p treated with first line EGFR TKIs and found that high expression of STAT3 and YAP were significantly correlated with shorter median progression-free survival (mPFS). mPFS was 9.6 months (m) (95% CI, 5.9 to 14.1) for p with low STAT3 and 18.4m (95% CI, 8.8 to 30.2) for p with high STAT3 mRNA expression (P Citation Format: Niki Karachaliou, Imane Chaib, Sara Pilotto, Jordi Codony, Xueting Cai, Xuefei Li, Ana Drozdowskyj, Carles Codony, Andrés Felipe Cardona, Guillermo López-Vivanco, Alain Vergnenègre, José Miguel Sánchez, Mariano Provencio, Filippo de Marinis, Enric Carcereny, Noemí Reguart, Rosario García-Campelo, Silvia Marin, Cristina Teixido, Isabella Sperduti, Sonia Rodríguez, Roger Estrada, Raimon Puig de la Bellacasa, José Luis Ramírez, Miguel Angel Molina-Vila, Caicun Zhou, Peng Cao, Patrick Ma, Trever Bivona, Rafael Rosell. Cotargeting EGFR, STAT3 and Src-Notch pathways: a promising approach to improve the efficacy of EGFR-TKIs in the treatment of NSCLC patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 265.

Backgroud: Tumor-derived circulating cell-free DNA (cfDNA) is a dynamic source for determination of tumor mutation status. We have previously demonstrated the prognostic value of BRAFV600 mutation status in pretreatment cfDNA (BRAF pre-cfDNA) in advanced melanoma patients (p) treated with BRAF inhibitors (median overall survival [OS] 7 months [m] vs 22m for BRAF pre-cfDNA positive and negative p, respectively p = 0.017)1. Based on these results, the Spanish Melanoma Group conducted a prospective study in 13 centers to determine the prognostic value of BRAFV600 mutation in pre-cfDNA, the change in mutation status at time of first evaluation (BRAF early-cfDNA), and the correlation of BRAF cfDNA dynamics with clinical evolution (GEM1304) (ClinicalTrials.gov Identifier: NCT01960634). Methods: One hundred and fifty nine plasma and serum samples from 66 stage IV BRAF mutant melanoma p were collected before and during treatment, until disease progression. A quantitative 5’-nuclease PCR based assay was used to determine BRAFV600 mutation status in cfDNA. Results: Most p were stage M1c (62%), treated with BRAF inhibitors (53%), and not previously treated (67%). BRAF pre-cfDNA was positive in 42 p (64%). Median OS was 6.4 m (95% CI: 10.9-23.6) and 17 m (95% CI: 3.5-9.2) for p with positive and negative BRAF pre-cfDNA, respectively (p = 0.06). Significant differences in OS were observed according to BRAF early-cfDNA negativization: 4.7 m (95%CI: 1.2-8.1) in those with persistence of BRAF in cfDNA (12 p), not reached (NR) in p with BRAF early-cfDNA negativization (11 p), and 22 m (95%CI:0.6-43.9) in those who continued to be negative (17 p) (p Conclusions: Patients with early negativization of BRAFV600 in cfDNA have excellent prognosis, at least as good as those with negative BRAF in pre-cfDNA. González-Cao et al. Mel Res 2015; 25:486 Citation Format: Maria Gonzalez Cao, Jose Luis Manzano, Virtudes Soriano, Teresa Puertolas, Ainara Soria, Clara Mayo, Margarita Magem, Miguel Angel Molina, Clara Montagut, Eva Muñoz, Delvys Rodriguez, Elizabeth Perez, Almudena Garcia, Javier Cortes, Nuria Jordana, Jordi Rodon, Niki Karachaliou, Rafael Rosell. BRAF mutation analysis in cell free tumoral DNA (cfDNA) of melanoma patients: results from the prospective study GEM1304 (Spanish Melanoma Group). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 468.

Introduction: The Epidermal Growth Factor Receptor (EGFR) signaling system is frequently unbalanced in non-small-cell lung cancer (NSCLC). Epidermal Growth Factor (EGF) could be an attractive target in these tumors. We used a “cancer vaccine” composed of human recombinant EGF conjugated to a carrier protein P64K. The vaccine is intended to induce antibodies against patients’ own EGF that would block EGF-EGFR interaction. Objectives: In NSCLC cell lines we assessed whether anti-EGF antibodies can inhibit the pathway activated by EGF-EGFR binding and whether sera from immunized patients can block downstream key signal transduction proteins. NSCLC cell lines were selected to determine whethergenetic alterations in EGFR, KRAS, ALK, BRAF or MET can have an impact on inhibition of EGFR pathway by anti-EGF antibodies. Experimental Procedures: Western blotting was used to evaluate the capacity of antibodies and sera from patients to inhibit EGFR activation pathway. MTT assays were performed to evaluate the effects on tumor cell proliferation. Results: The anti-EGF antibodies showed a dose-dependent inhibitory effect on EGF-induced activation of EGFR and downstream pathway molecules such as Akt and ERK 1/2. This inhibition was observed in all cell lines tested, including KRAS, EGFR and BRAF mutated cells as well as cells with ALK translocations or MET amplification. Sera from 10 patients immunized with the P64K-EGF vaccine were then tested. All sera reversed EGF-induced activation of EGFR and ERK 1/2, but some sera were significantly more potent than others. By contrast, two control sera from non-vaccinated patients failed to inhibit EGF-induced phosphorylation of EGFR and ERK 1/2. Finally, anti-EGF antibodies were also able to block the stimulatory effects of EGF on the growth of some NSCLC cell lines. Conclusions: Anti EGF antibodies and sera from patients immunized with an “EGF vaccine”, called otherwise EGF Pathway Targeted Immunization inhibits EGFR signaling in NSCLC cell line models. This effect is independent of cells’ genetic background. Citation Format: Jordi Codony-Servat, Miguel Angel Molina-Vila, Jordi Bertran-Alamillo, Rafael Rosell, Erik D’Hondt. Inhibition of epidermal growth factor receptor pathway by epidermal growth factor antibodies in non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2368.

Background: Targetable rearrangements in anaplastic lymphoma kinase (ALK), ROS1, and RET genes are present in approximately 7% of patients (p) with advanced NSCLC. Current methods for detecting gene fusions are based on FISH (FDA-approved companion diagnostic test for ALK), immunohistochemistry (IHC) or RT-PCR. However, these tests have disadvantages in terms of sensitivity, cost and throughput, and often show discrepancies. The nCounter platform allows simultaneous detection with no enzymatic reaction, within 72 hours, of several fusion genes in formalin-fixed paraffin-embedded (FFPE) samples using a transcript-based method. Methods: A custom set of 5′and 3′ probes and fusion-specific probes to detect ALK, ROS1 and RET fusion transcripts was designed and evaluated. A panel of ALK-ROS1-RET positive cell lines (H2228, H3122 [EML4-ALK], SUDHL-1 [NPM-ALK], HCC78 [SLC34A2-ROS1], BaF3 pBABE [CD74-ROS1], LC2/ad [RET]) and negative cell lines (PC9, H1975 [EGFR mut], H460, H23 [KRAS mut]) was used to validate the technique. Then, a total of 70 FFPE samples was analyzed, 49 of them positive by FISH, IHC and/or qRT-PCR for ALK (n = 30), ROS1 (n = 17) and RET (n = 2). Total RNA was isolated from cell lines and FFPE and < 225 ng were used for hybridization. Raw counts were normalized using positive controls, negative controls and four house-keeping genes (GAPDH, GUSB, OAZ1 and POLR2A) as described in Lira et al. J Mol Diagn 2013. Positive and negative translocations were defined by two criteria: (1) a 3’/5’ ratio score of > 2.0 and ≤ 2.0 respectively or/and (2) a signal for a fusion-specific probe above background. Response to crizotinib by RECIST criteria was retrospectively collected in p with ALK-positive NSCLC by any technique. Results: nCounter sensitivity to detect fusion transcripts ALK, ROS1 and RET in cell lines was 100% using the two criteria (3’/5’ and direct probes) and specificity was also 100%. Among 20 ALK-FISH-positive p, ALK 3’-5’ scoring was positive in 18 (95%). One p was non-evaluable (NE) by ALK 3’-5’ scoring. Among 48 ALK-FISH-negative p, nCounter score was positive in 13 (27%). All p positive for ALK by nCounter were either positive or NE for ALK by IHC. A total of 17 p were treated with crizotinib, 16 of whom responded to treatment and were positive by nCounter. Regarding FISH, five p responding to crizotinib were negative and one was NE. Finally, one p not responding to crizotinib was positive by RT-PCR but negative by nCounter. Conclusion: The ALK/ROS1/RET nCounter-based assay is a highly sensitive screening assay that identifies ALK-FISH-negative/NE NSCLC patients who could benefit from treatment with ALK inhibitors. Citation Format: Cristina Teixido, Noemí Reguart, Ana Giménez-Capitán, Miguel Ángel Molina-Vila, Patricia Galván, Sonia Rodriguez, Laia Paré, Santiago Viteri, Vicente Peg, Zaira Yeste, Núria Viñolas, Rafael Rosell, Aleix Prat. Comparison of nCounter, immunohistochemistry, RT-PCR and FISH to detect ALK, ROS1 and RET rearrangements in advanced non-small cell lung cancer (NSCLC). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4344.

The BELIEF trial (NCT01562028) examined the efficacy of E+B for the 1st line treatment of European p with advanced NSCLC harboring exon 19 deletion or exon 21 L858R epidermal growth factor receptor (EGFR) mutations with or without pretreatment T790M. Median progression-free survival (mPFS) was 13.8 months (m) (95% CI 10.3-16.2) for the 109 p enrolled in the study. mPFS was 16m (95% CI 13.1-not estimable [NE]) and 10.4m (95% CI 9.2-15.6) for the 37 T790M(+) p and 72 T790M(-) p, respectively (P = 0.089). We have previously shown that low BRCA1 levels neutralize the negative effect of pretreatment T790M and are associated with longer PFS to erlotinib. Low astrocyte elevated gene 1 (AEG1) levels are associated with longer PFS to erlotinib. A secondary objective of the BELIEF trial was the assessment of the prognostic impact of BRCA1 and AEG1 mRNA expression in the baseline tumor tissue. We assessed the baseline mRNA expression levels of BRCA1 and AEG1 and correlated them with PFS and response in the 109 EGFR-mutant NSCLC p of the BELIEF trial. BRCA1 and AEG1 mRNA levels were estimable by quantitative-real time PCR in 46 and 61 p respectively. Expression levels were divided into two groups according to their median. No statistically significant associations were found among the expression of the two biomarkers and gender, smoking status, histology, PS or type of EGFR mutation. The association of BRCA1 and AEG1 mRNA with PFS did not differ according to T790M status (interaction P = 0.81 and 0.42, respectively). Among the T790M (+) p, those with low BRCA1 mRNA had a longer mPFS of 24.6m (95% CI: 4.9-NE) compared to 15.4 m (95% CI: 2.7-NE) for p with high BRCA1 mRNA; P = 0.63. For all and T790M (-) p, BRCA1 levels did not differentiate mPFS to E+B (low vs high BRCA1 mRNA, 11.0 m [95% CI: 6.0-NE] vs 9.5 m [95% CI: 7.2-NE]; P = 0.99 and 6.9 m [95% CI: 2.1-NE] vs 9.4 m [95% CI: 4.1-NE]; P = 0.71, respectively). Similarly, among the T790M (+) p, those with low AEG1 expression had a longer mPFS of 24.6m (95% CI: 4.9-NE) compared to 15.4 m (95% CI: 5.2-NE) for those with high AEG1 mRNA; P = 0.93. For all and T790M(-) p, AEG1 levels did not differentiate mPFS to E+B (low vs high AEG1 mRNA, 10.3 m [95% CI: 8.4-24.6] vs 15.4 m [95% CI: 6.0-33.9]; P = 0.90 and 10.3m [95% CI: 8.4-NE] vs 13.3m [95% CI: 4.7-33.9] P = 0.67). No statistically or clinically significant associations were found among BRCA1, AEG1 mRNA levels and response to E+B. The BELIEF trial reconfirms our previous findings that pretreatment EGFR T790M is present in 34% of the patients. E+B is associated with a prolonged mPFS of 24.6m for EGFR-mutant p with both pretreatment EGFR T790M and low BRCA1 mRNA expression, as observed in our previous study with erlotinib alone. Similar results were observed for AEG1 levels though the interaction was not statistically significant for either biomarker. Citation Format: Rafael Rosell, Niki Karachaliou, Ana Giménez-Capitán, Carles Codony-Servat, Oliver Gautschi, Enriqueta Felip, Alessandra Curioni-Fontecedro, Solange Peters, Santiago Ponce-Aix, Martin Früh, Miklos Pless, Sanjay Popat, Sinead Cuffe, Paolo Bidoli, Adolfo Favaretto, Roswitha Kammler, Urania Dafni, Zoi Tsourti, Miguel Angel Molina-Vila, Rolf A. Stahel. The role of BRCA1 and AEG1 mRNA expression in advanced non-small-cell lung cancer (NSCLC) patients (p) with EGFR activating and pretreatment T790M mutations receiving the combination of erlotinib plus bevacizumab (E+B) in the BELIEF trial. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 269.

Background: Aberrant activity of MET, FGFR1 and AXL has been associated with development of resistance to EGFR inhibitors in EGFR-mutated non-small cell lung cancer (NSCLC) patients. S49076 is a potent ATP-competitive tyrosine kinase inhibitor (TKI) of MET, AXL/MER, FGFR1/2/3 currently in phase I clinical trials. Methods: We obtained six resistant lines by treating EGFR-mutated, TKI-sensitive PC9 cells with increasing concentrations of gefitinib (GR1-5) or erlotinib (ER). The six resistant cell lines conserved the exon 19 deletion but the T790M resistance mutation only emerged in two (GR1, GR4), which remained sensitive to AZD9291, a third generation EGFR TKI. Six new AZD9291-resistant cell lines were derived from GR1 and GR4 by exposure to increasing concentrations of the inhibitor. Three of the AZD9291 resistant cell lines lost the exon 19 and T790M mutations, one conserved only the sensitizing mutation and two kept both mutations but with the T790M dropping to very low allelic fractions (1% and 0.03%). All resistant cell lines were characterized for AXL, MET, MER, FGFR1 and FGFR2 expression by Q-RT-PCR, immunohistochemistry and Western blotting. The effects of S49076 on the parental and resistant cell lines were analyzed by MTT, colony formation, basal membrane invasion and migration assays. Western blotting of key signal transduction proteins was used to gain insight in the drug's mechanism of action. Results: AXL overexpression was the most common event related to gefitinib/erlotinib resistance in the panel of cell lines, with T790M mutation, FGFR1 or BCL-2 overexpression and MET activation being less frequent events. Regarding AZD9291 resistance, AXL upregulation was again widespread, with loss of EGFR mutations as the second most frequent event. In proliferation assays, S49076 showed strong antitumor activity against all PC9-derived cell lines with acquired resistance to EGFR TKIs (erlotinib, gefitinib or AZD9291) and not carrying the T790M mutation, with IC50 of 0.2-1.2 μM and less than 10% surviving cells at 1 μM of drug in most cases. In contrast, S49076 was less active against the parental PC9 cells and the T790M positive resistant lines, with more than 50% of surviving cells at 50 μM of drug. S49076 also inhibited the anchorage-independent growth and migration of the resistant cell lines. A correlation was found between mRNA and protein levels of AXL and sensitivity to S49076. Also, PARP cleavage and moderate but reproducible inhibition of AKT phosphorylation by S49076 were observed exclusively in the non-T790M resistant cell lines. Conclusions: S49076 shows strong activity in preclinical assays against EGFR mutated cell lines resistant to EGFR TKIs not harboring EGFR T790M mutation and overexpressing AXL. A clinical trial of S49076 in non-T790M patients progressing to EGFR TKI and overexpressing AXL or MET has been initiated. Citation Format: Jordi Bertran-Alamillo, Miguel A. Molina, Cattan Valérie, Mike Burbridge, Cristina Teixido, Jordi Codony-Servat, Carles Codony-Servat, Rafael Rosell. S49076, a kinase inhibitor of AXL, MET and FGFR with strong, selective preclinical activity against tumor cells with acquired resistance to EGFR inhibitors not carrying the T790M mutation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 336.

The inhibition of carboxypeptidase A (CPA) by its natural inhibitor from potato (PCI) has been widely analysed with theoretical and experimental methods. Several mutants of PCI have been obtained in order to study the physico-chemical properties related to the inhibition. Point mutations were performed in the C-tail of PCI given its fundamental role in the inhibition. The inhibition constant and the dissociation free energy of the complexes PCI-CPA was experimentally obtained for each mutant. The mutants were divided in two sets, those where the mutation was intrinsically affecting the conformation of the PCI C-tail, and those where the mutation affected the interaction between PCI and CPA. The crystallographic structure of PCI, as found in its complex with bovine carboxypeptidase A, was used to model the structure of these mutants.

11581Background: Targeted therapy has improved outcomes in EGFR mutated and ALK translocated NSCLC but options for squamous cell carcinoma and KRAS-mutated tumors remain scarce. Genome re-replicati...

e23039Background: One of the most promising developments in translational cancer has been the emergence of liquid biopsy as a non-invasive biomarker. CTCs and cfDNA offer valuable prognostic and pr...

It is widely acknowledged that there is a need for molecular profiling in non-small-cell lung cancer. For example, treatment based on EGFR mutation status has attained successful results. However, in spite of excellent initial response to oral EGFR tyrosine kinase inhibitors (TKIs), progression-free survival is still limited. Current research has focused mostly on acquired resistance mechanisms, such as overexpression of AXL and loss of the Mediator MED12. In this review, in contrast, we discuss adaptive, rather than acquired, resistance. Adaptive resistance can occur almost immediately after starting targeted therapy through a rapid rewiring of cancer cell signaling. By losing ERK negative feedback on receptor tyrosine kinase (RTK) expression, cancer cells are exposed to the stimuli of several ligands, and the ensuing activation of several RTKs reprograms all the canonical signaling pathways. The overexpression of several RTKs was observed in breast cancer cell lines treated with a MEK inhibitor and in BRAF(V600E) melanoma cell lines treated with BRAF inhibitors. This rebound effect of overexpression of several RTKs, including ERBB3, also occurs in lung cancers driven by Kras or EGFR mutations when treated with MEK, PI3K or dual PI3K/mTOR inhibitors. Synthetic lethality can be effectively induced by co-targeting these overexpressed RTKs. We speculate that in patients with EGFR mutations, adaptive resistance occurs in a significant proportion of patients. Rebiopsies performed hours after starting treatment with EGFR TKIs can identify which RTKs are overexpressed after treatment. Efficient co-targeting of these RTKs can induce synthetic lethality and help overcome the limited effect of EGFR TKI monotherapy.

7522 Background: When EGFR compound mutants (exon 19 deletions or L858R plusT790M) are present at the time of clinical resistance to erlotinib, patients attain longer overall survival (OS) (Oxnard et al. CCR 2011). The H1975 cell line, harboring L858R plus T790M prior to treatment, is sensitive to gefitinib (Sordella et al. Science 2004; Faber et al. Cancer Discovery 2011), though the compound mutant may become dominant after multiple passages in vitro, leading to resistance (Pao et al. PLoS Med 2005). Methods: The EURTAC trial (clinicaltrials.gov NCT00446225) randomized 174 p with EGFR exon 19 deletions or L858R mutations to receive erlotinib or chemotherapy. Progression-free survival (PFS) was 9.7 months (m) vs 5.2 m, respectively (P

In order to optimize the production of recombinant potato carboxypeptidase inhibitor (rePCI), a protein with 39 amino acid residues and three disulphide bridges, by Escherichia coli MC1061[pIMAM3], the effects of various parameters were investigated. Production of rePCI in M9CAS medium was optimal at 37°C and using low concentrations of glycerol as a carbon source. Increasing concentrations of glycerol caused a decrease in the production of rePCI, which was almost totally inhibited above 1% glycerol. Relatively high concentrations of oligoelements in the culture medium also inhibited the production of rePCI. We obtained a 100-fold increase in the production of rePCI, from 2 to 200 mg/l, when growing bacteria in a fed-batch aerobic culture using a 2-1 fermentor. RePCI was found exclusively in the supernatant, although the genetic construction was designed for it to be released into the periplasmic space. Large quantities of rePCI could be easily purified from the supernatants of these cultures. Our conditions of fed-batch, aerobic fermentation could be used for overproduction to high levels of other recombinant proteins.

The present article summarizes the discussions of the 3rd European Science Foundation–University of Barcelona (ESF–UB) Conference in Biomedicine on Pharmacogenetics and Pharmacogenomics, which was held in June 2010 in Spain. It was focused on practical applications in routine medical practice. We provide practical recommendations for ten different clinical situations, that have either been approved or not approved by regulatory agencies. We propose some comments that might accompany the results of these tests, indicating the best drug and doses to be prescribed. The discussed examples include KRAS, cetuximab, panitumumab, EGFR–gefitinib, CYP2D6–tamoxifen, TPMT–azathioprine–6-mercaptopurine, VKORC1/CYP2C9–warfarin, CYP2C19–clopidogrel, HLA-B*5701–abacavir, HLA-B*5701–flucloxacillin, SLCO1B1–statins and CYP3A5–tacrolimus. We hope that these practical recommendations will help physicians, biologists, scientists and other healthcare professionals to prescribe, perform and interpret these genetic tests.

The potential differential effect of first-line treatment and molecular mechanisms on survival to second-line chemotherapy or EGFR tyrosine kinase inhibitors (TKIs) in non-small-cell lung cancer (NSCLC) has not been fully investigated. In particular, CHFR is frequently methylated in NSCLC and may influence outcome. We analyzed the outcome of second-line chemotherapy or EGFR TKIs in 179 of 366 patients who had been treated in an ERCC1 mRNA-based customized cisplatin trial and correlated the results with CHFR methylation status. CHFR methylation in circulating DNA was examined by methylation-specific assay. A panel of seven human EGFR wild-type NSCLC cell lines was characterized for their sensitivity to sequential treatment with cisplatin and erlotinib, and the results were correlated with CHFR. Patients who had received first-line docetaxel/cisplatin attained an overall survival of 19.2 months when treated with second-line EGFR TKIs, in comparison with 10.7 months when treated with second-line chemotherapy (P = 0.0002). However, for patients who had received first-line docetaxel/gemcitabine, overall survival was 14.8 months with EGFR TKIs and 10.8 months with chemotherapy (P = 0.29). For patients with unmethylated CHFR overall survival to EGFR TKIs was 21.4 months, and 11.2 months for those with treated with chemotherapy (P = 0.0001). In the only lung tumor cell line not expressing CHFR, pretreatment with cisplatin was antagonistic to erlotinib, while it was synergistic in the other six lines. Second-line EGFR TKIs improved survival in patients receiving first-line cisplatin-based treatment. Unmethylated CHFR predicts increased survival to EGFR TKIs.

Background Metastatic non-small-cell lung cancer (NSCLC) has a dismal prognosis. EGFR is overexpressed or mutated in a large proportion of cases. Downstream components of the EGFR pathway and crosstalk with the NF-κB pathway have not been examined at the clinical level. We explored the prognostic significance of the mRNA expression of nine genes in the EGFR and NF-κB pathways and of BRCA1 and RAP80 in patients in whom EGFR and K-ras gene status had previously been determined. In addition, NFKBIA and DUSP22 gene status was also determined. Methods mRNA expression of the eleven genes was determined by QPCR in 60 metastatic NSCLC patients and in nine lung cancer cell lines. Exon 3 of NFKBIA and exon 6 of DUSP22 were analyzed by direct sequencing. Results were correlated with outcome to platinum-based chemotherapy in patients with wild-type EGFR and to erlotinib in those with EGFR mutations. Results BRCA1 mRNA expression was correlated with EZH2, AEG-1, Musashi-2, CYLD and TRAF6 expression. In patients with low levels of both BRCA1 and AEG-1, PFS was 13.02 months, compared to 5.4 months in those with high levels of both genes and 7.7 months for those with other combinations (P = 0.025). The multivariate analysis for PFS confirmed the prognostic role of high BRCA1/AEG-1 expression (HR, 3.1; P = 0.01). Neither NFKBIA nor DUSP22 mutations were found in any of the tumour samples or cell lines. Conclusions The present study provides a better understanding of the behaviour of metastatic NSCLC and identifies the combination of BRCA1 and AEG-1 expression as a potential prognostic model.

Activating mutations in the form of deletions in exon 19 (del 19) or the missense mutation L858R in the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) predict outcome to use of EGFR tyrosine kinase inhibitors (TKIs), such as gefitinib and erlotinib. Pooled data from several phase II studies show that gefitinib and erlotinib induce responses in over 70% of NSCLC patients harboring EGFR mutations, with progression-free survival (PFS) ranging from 9 to 13 months. Two studies in Caucasian and Asian patients have confirmed that these subgroups of patients attain PFS up to 14 months. These landmark outcomes have been accompanied by new challenges, primarily the additional role of chemotherapy and the management of tumors with the secondary T790M mutation that confers resistance to EGFR TKIs. Mechanisms of resistance to reversible EGFR TKIs should be further clarified and could be related to modifications in DNA repair.

Background Immunohistochemistry (IHC) with mutation-specific antibodies may be an ancillary method of detecting EGFR mutations in lung cancer patients. Methods EGFR mutation status was analyzed by DNA assays, and compared with IHC results in five non-small-cell lung cancer (NSCLC) cell lines and tumor samples from 78 stage IV NSCLC patients. Results IHC correctly identified del 19 in the H1650 and PC9 cell lines, L858R in H1975, and wild-type EGFR in H460 and A549, as well as wild-type EGFR in tumor samples from 22 patients. IHC with the mAb against EGFR with del 19 was highly positive for the protein in all 17 patients with a 15-bp (ELREA) deletion in exon 19, whereas in patients with other deletions, IHC was weakly positive in 3 cases and negative in 9 cases. IHC with the mAb against the L858R mutation showed high positivity for the protein in 25/27 (93%) patients with exon 21 EGFR mutations (all with L858R) but did not identify the L861Q mutation in the remaining two patients. Conclusions IHC with mutation-specific mAbs against EGFR is a promising method for detecting EGFR mutations in NSCLC patients. However these mAbs should be validated with additional studies to clarify their possible role in routine clinical practice for screening EGFR mutations in NSCLC patients.

2-[(Triphenylphosphoranylidene)amino]benzyl alcohol 2 reacts with isocyanates or isothiocyanates in refluxing toluene to give the corresponding 2-alkyl(aryl)amino-4H-3,1-benzoxazines 5. Iminophosphorane 2 also reacts with carbon dioxide and carbon disulfide to give 4H-3,1-benzoxazin-2(1H)-one (6) and -thionine (7), respectively

The World Space Observatory – Ultraviolet (WSO-UV) is an international space observatory for observation in UV spectral range (100 350 nm), that is beyond the reach of ground-based instruments but where most of astrophysical processes can be efficiently studied with unprecedented capability. The WSO-UV has been driven by the needs of scientists to have an UV facility in the horizon of the next decade. It will be the only two meters class mission for UV observations in the postHubble Space Telescope era. The WSO-UV project is managed by a large, world-wide, consortium lead by the Federal Space Agency (Roscosmos, Russia). The WSO-UV consists of a 1.7 m aperture telescope (under responsibility of Russia) with instrumentation designed to carry out high resolution spectroscopy (spectral resolution about 55000), long-slit low resolution spectroscopy (spectral resolution about 2000) and direct sky imaging. The WSO-UV Ground Segment (GS) is under development by Spain and Russia and both countries will coordinate the Mission and Scientific operations and will provide the satellite tracking stations for the project. The nominal lifetime is 5 years with a planned extension to 10 years. The project launch date set in late 2013. Aiming to maximize the scientific return along such a long lifetime, the ground segment architecture is to be based in a modular approach, relying in a common framework able to run together different subsystems developed from different agencies and institutes, which may be fully upgraded and even replaced along the years. The WSO-UV GS is comprised of all the infrastructure and facilities involved in the preparation and execution of the WSO-UV mission operations, which typically encompass real-time monitoring and control of the spacecraft, telescope and instruments as well as reception, processing and storage of the scientific data. In principle, there will be two complete ground segment systems, the Russian one will be located in Moscow (Lavochkin Association and Institute of astronomy RAS), and the Spanish one will be sited at Madrid (Universidad Complutense de Madrid, UCM). The satellite operations will be shared between both Ground Control Centers transferring the mission control from one center to the other on a regular basis. The Ground Segment development is based as much as possible on GMV’s existing products for telemetry and telecommand systems, flight dynamics and science processing. One of the main challenges on GS development is the management of shared information between both centers and the alignment of all the operational data (telemetry, telecommand and planning) according to the operational shifts. The WSO-UV Context Manager component will be in charge of the synchronization of the Ground Control Centers at different levels and avoiding operational conflicts. Data received in either ground station will be sent via a fast, secure data link to the two mission control, but only one will be able to command the spacecraft at every time. Current GS scenario is based on real-time reaction to target of opportunity (ToO) alerts. This implies creation of new timeline of commands, interruption of current one and starting of the new one, which will target the spacecraft

Background: Activating mutations in the epidermal growth factor receptor gene (EGFR) confer hypersensitivity to the tyrosine kinase inhibitors gefitinib and erlotinib in patients with advanced non-small-cell lung cancer. We evaluated the feasibility of large-scale screening for EGFR mutations in such patients and analyzed the association between the mutations and the outcome of erlotinib treatment. Methods: From April 2005 through November 2008, lung cancers from 2105 patients in 129 institutions in Spain were screened for EGFR mutations. The analysis was performed in a central laboratory. Patients with tumors carrying EGFR mutations were eligible for erlotinib treatment. Results: EGFR mutations were found in 350 of 2105 patients (16.6%). Mutations were more frequent in women (69.7%), in patients who had never smoked (66.6%), and in those with adenocarcinomas (80.9%) (P

Histiocytic sarcoma (HS) is a rare malignant neoplasm of the lymphohematopoietic system, that occurs in lymph nodes, skin and at extranodal sites, particularly the gastrointestinal tract. Although it shows characteristics histological and immunohistochemical features, it may be misdiagnosed. We report a 67 year-old female patient presenting with colicky abdominal pain and vomiting. A CT scan of the abdomen revealed a tumor in the ileum, that was surgically removed. On pathology, the neoplastic cells displayed large abundant eosinophilic cytoplasm, with bizarre-shaped nuclei, that expressed CD 45, CD 68 and lisozyme. The diagnosis of HS requires the use of a panel of immunohistochemical markers and may be supported by ultrastructural findings.

Histiocytic sarcoma (HS) is a rare malignant neoplasm of the lymphohematopoietic system, that occurs in lymph nodes, skin and at extranodal sites, particularly the gastrointestinal tract. Although it shows characteristics histoiogical and immunohistochemical features, it may be misdiagnosed. We report a 67 year-old female patient presenting with colicky abdominal pain and vomiting. A CT sean of the abdomen revealed a tumor in the ileum, that was surgically removed. On pathology, the neoplastic cells displayed large abundant eosinophilic cytoplasm, with bizarre-shaped nuclei, that expressed CD 45, CD 68 and ¡isozyme. The diagnosis of HS requires the use of a panel of immunohistochemical markers and may be supported by ultrastructural findings .

ERBB2, a ligand-less membrane receptor, is frequently overexpressed in a number of human tumors, contributing to uncontrolled cell proliferation. In some cases, gene amplification correlates with protein overexpression and predicts response to trastuzumab. We analyzed the expression of ERBB2 in a group of 40 patients diagnosed with infiltrating squamous cervical carcinomas (ISCC) using a microarray. Immunochemistry was performed using two different antibodies, one against the extramembrane domain and the other one for the intramembrane domain. Ten of the 40 cases included in the study could not be evaluated. Of the 30 remaining biopsies, 13 (42%) showed immunoreactivity only with the antibody against the intramembrane domain. In 5 (16.12%), both intramembrane and extramembrane immunoreactivity was observed, and 12 (40%) were negative for both antibodies. Looking at our results, we propose that, in some ISCC, there is a rupture of the ERBB2 receptor, and this event, with slight genetic amplification, could explain the unfavorable response to trastuzumab observed in some ISCC descript for some authors.

HISPASAT Group is operating now four Satellites:1C, 1D, and SPAINSAT (to cover the demand of governmental services for the Spanish Ministry of Defense) in 30o West and Amazonas-1 in 61o West. In preparation to cover the emerging demand in 61o West, HISPASAT has under construction Amazonas-2 satellite, to be placed in 61o West along year 2009. Additional satellites are under study for both orbital positions in the plan to increase the available on orbit capacity. HISPASAT manages the Satellite Control Operations for Telemetry and Telecommand functions by using platform specific Satellite Control Centers (SCC), therefore having at the present stage three different SCC platfomrs. This approach is being revisited and HISPASAT’s philosophy is now oriented towards a more flexible and easy to operate concept that implies to integrate the satellite operations using a common control platform. That continues the trend already initiated with the Flight Dynamics with GMV’s focusGEO and will be complemented in the Satellite Control Center area with GMV’s hifly®.

In spite of the dismal outcome of glioblastoma multiforme (GBM), we are in a position to provide a ray of hope to patients and families. Methylation of MGMT in tumor occurs in approximately a third of patients and predicts meaningful response and survival to adjuvant radiotherapy plus temozolomide. Limited access to tumor tissue in some patients could be circumvented by examining MGMT methylation in circulating serum DNA, although this approach needs to be validated. Molecular signatures are also promising prognostic and predictive markers, and clinical trials should be carried out to validate their use in the selection of patients for specific targeted therapies. Gene expression by quantitative PCR of key components of these molecular signatures could pave the way for easy identification of different subgroups of patients. Translational clinical trials are warranted in order to detect the subgroups of patients resistant to radiotherapy who may derive benefit from novel therapies, including antiangiogenic drugs.

Metastatic stage IV non-small-cell lung cancer (NSCLC) has a grim outcome. The median survival does not exceed 11 months, with no differences according to different cisplatin-based regimens. However, this clinical observation is spurious from the molecular point of view, bewcausse DNA repair genes involved in several pathways act as differential modulators of chemosensitivity. Low expression of ERCC1 or BRCA1 predicts higher sensitivity to cisplatin and other DNA-damaging agents, such as etoposide and mitomycin, while conferring resistance to antimicrotubule drugs, such as vinblastine and paclitaxel. In contrast, high expression of these genes confers resistance to cisplatin and sensitivity to antimicrotubules.

e19085 Background: Repeat biopsies of cancer p are useful for monitoring molecular evolution of the disease and the possible appearance or disappearance of key somatic mutations, particularly those...

e13516 Background: Targeted lung cancer therapy has undoubtedly made a difference to the treatment of EGFR mutation and ALK translocation carriers. However, targeted therapies for other subgroups l...

1042 Background: Neoadjuvant chemotherapy is generally used for treatment of both early stage and locally advanced breast cancer. However, robust prognostic or predictive biomarkers are needed in t...

The provision of closed solution for flight dynamics applications has increased its demand in the last years. Since 2000, focusSuite has tried to provide reliable and compact solutions for flight dynamics in several contexts and mission profiles. focusSuite was conceived as a software package that should compile software and knowledge in the flight dynamics and operations areas. A deep survey of requirements and flight dynamics operator needs was performed in the initial stages of its development. The conclusion was a first approach to focusSuite that finalised with the implementation of focusGEO to support geostationary flight dynamics operations for EUTELSAT and HISPASAT in 2002. The evolution of focusSuite has been driven by two main sources of input: GMV’s internal R+D activities searching for solutions more adequate to new project needs and also by specific requirements from operators and analysts, The result is the consolidation of focusSuite as platform for GMV’s flight dynamics solutions. In 2004, ESA’s flight dynamics software package is adapted and integrated into focusSuite to become the solution for the GALILEO test bed GSTB-V2; this is the first realisation of focusLEO. In parallel, this version is used to prototype the integration of the EPS flight dynamics system to proof the feasibility for Earth observation missions. In parallel to integration of the operational systems, several auxiliary components are developed to fulfil specific needs in different areas around flight dynamics: focusASPECT For telemetry processing and AOCS monitoring, Visualfocus for 2D/3D visualisation, focusCloseAp for collision risk assessment, focusLEOP to support the Launch and Early Orbit Phase for geostationary satellites, to mention some. Today and in the coming future, focusSuite is supporting and will further support the flight dynamics operations of a long list of satellites in the commercial market. Additionally, design versatility is being proven adapting focusCn to the very demanding requirements of GALILEO. This paper addresses all the experience and lessons learned from the evolution of focusSuite and the adaptation of new and already operational flight dynamics systems into this infrastructure.

This article adds the Spanish grid code to the previously published works about the comparison of international regulations for connection of wind turbines to the network. Specifically, is focused on the reactive power generated by wind turbines when they are submitted to a voltage dip. Taken advantage of the voltage dips field tests carried out to the GAMESA G80 wind turbines, several definitions of reactive power are calculated and the results obtained for two representative voltage dip tests are presented: a three-phase and a phase-to-phase voltage dip

The authors would like to thank to the Ministerio de Eduacion of Spanish Government for supporting this research under the Project DPI2001-2779-C02-01, as well to thank the European Union for their economic support.