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Cancer is one of the most important health problems in the world, affecting a large part of the population. Although there are different treatments such as surgery, radiotherapy or chemotherapy, the cancer currently has no cure, in the strict sense of the word. The numbers of those affected continue to increase, and the future forecasts are not very comforting. Cancer occurs due to alterations in the genes involved in cell growth, survival and propagation, resulting in a set of common characteristics. These characteristics are known as the hallmarks of cancer, and allow to organize and rationalize this complex disease. ErbB (erythroblastic) receptors play a very important role, they’re essential mediators of cell proliferation and differentiation in the development of the embryo and adult tissues, and their inappropriate activation is associated with the development and severity of the cancer. Members of this tyrosine kinase receptor family are: EGFR (ErbB1 or HER1), HER2 (Neu or ErbB2), HER3 (ErbB3) and HER4 (ErbB4). EGFR and HER2 are associated with cancer, since its alteration is found in most cancers. Recently, it has been seen that HER3 receptor also has an important role, in addition, it can act as a new target to treat this disease. Each of these receptors have several ligands with which they bind to be activated, with the exception of HER2 that isn’t known any ligand. Once activated, the receptors dimerize: homodimerization (among them) or heterodimerization (between family members). The heterodimers formed by HER2 and HER3 form the most robust signalling complexes of the ErbB family. One of the characteristics of HER3 is its unique and powerful ability to activate the downstream signalling path PI3K and Akt. It’s key to the control of many biological processes critical for tumorigenesis. In addition, HER3 is also associated with cancer resistance, which is the main problem of current therapies. With these evidences emphasizing the importance of HRE3 signalling in human cancers and their resistance, today there are more than 40 different therapeutic agents that are being tested targeting HER3, in addition to combinatorial therapies that act against different combinations of receptors

Poly(ADP-ribosyl)ation (PARylation) is a widespread and highly conserved post-translational modification catalysed by a large family of enzymes called poly(ADP-ribose) polymerases (PARPs). PARylation plays an essential role in various cardinal processes of cellular physiology and recent approvals and breakthrough therapy designations for PARP inhibitors in cancer therapy have sparked great interest in pharmacological targeting of PARP proteins. Although, many PARP inhibitors have been developed, existing compounds display promiscuous inhibition across the PARP superfamily which could lead to unwanted off-target effects. Thus the prospect of isoform-selective inhibition is being increasingly explored and research is now focusing on understanding specific roles of PARP family members. PARP-2, alongside PARP-1 and PARP-3 are the only known DNA damage-dependent PARPs and play critical roles in the DNA damage response, DNA metabolism and chromatin architecture. However, growing evidence shows that PARP-2 plays specific and diverse regulatory roles in cellular physiology, ranging from genomic stability and epigenetics to proliferative signalling and inflammation. The emerging network of PARP-2 target proteins has uncovered wide-ranging functions of the molecule in many cellular processes commonly dysregulated in carcinogenesis. Here, we review novel PARP-2-specific functions in the hallmarks of cancer and consider the implications for the development of isoform-selective inhibitors in chemotherapy. By considering the roles of PARP-2 through the lens of tumorigenesis, we propose PARP-2-selective inhibition as a potentially multipronged attack on cancer physiology. The JY laboratory is supported by grants from Spanish Ministerio de Economía y Competitividad (SAF2014-53467-R), Fundació La Marató de TV3 (20134130), and CIBERehd.

We aimed to assess changes in the total effective xenoestrogen burden (TEXB) -a biomarker of combined effect of mixtures of xenoestrogens- in breast cancer patients at surgery (breast adipose tissue) and at different time points during an 18-month follow-up (abdominal adipose tissue), and to analyze the potential influence of socio-demographic, reproductive, tumor, and treatment characteristics on TEXB levels. TEXB-alpha (due to persistent organohalogenated chemicals) and TEXB-beta (due mainly to endogenous estrogens) were quantified in 44 women. TEXB values significantly increased over follow-up (p

Cancer can be defined as a family of complex disease characterized by a cellular hyperproliferation and the capacity of the cell to invade surrounding tissue and metastasize to other organs in the body. The development and progression of cancer is a complex multistep process in which cells gradually acquire mutations altering the cellular mechanisms until the cell becomes malignant. Cancer develops and progresses through the acquisition of the above mutations or alterations across biological levels. Downstream of these alterations are expression changes in many genes at each stage of the disease. Sets of genes (also called signatures whose differential expression or profiles have prognostic or predictive (in terms of prediction of drug-response) values have been identified for almost every type of cancer. In some cases, several signatures have proved to be useful in independent evaluations, although, intriguingly, their overlap in gene identities was minimal. Then, integrative approaches using different types of gene and protein relationships have demonstrated the existence of biological convergence among apparently disparate gene sets. Moreover, integrating data from he network of known protein-protein interactions (hereafter interactome network) has been shown to improve the reproducibility and accuracy of prognostic signatures. However, in this scenario, the network topological patterns linked to the dynamic molecular alterations that characterize cancer development and progression, and treatment response, remain unknown. Several key genetic, transcriptomic and molecular determinants of cancer therapeutic response have been identified in recent years. Specific genetic alterations have been demonstrated to mediate the existence and/or promote the acquisition of therapeutic resistance. In addition, sets of transcripts have been identified whose profiles have predictive value for prognosis and/or treatment benefit. Successively, the integration and modeling of data from different types of molecular interactions has been shown to enhance understanding of the mechanisms of cancer progression and therapy response. In this scenario, however, cancer patients all too frequently show no or only modest benefit from a given therapy. The persistence of this fundamental clinical problem has been partially attributed to the lack of specific biomarkers. However, the identification of a comprehensive measure of cancer cell activity may support the interpretation of therapy efficacy. At a cancer system level, several studies have shown extensive molecular rewiring and increased signaling entropy, which may endorse the characteristic robustness of cancer. Given these observations, an a priori understanding of therapeutic response should be supported by an integrated measure of cancer network activity. In this study, firstly, we hypothesized that the features of dynamism and robustness intrinsic to cancer should also be present at different biological levels and, in particular, evident within the topology of he interactome network. To assess this hypothesis we analyzed the impact of cancer-related expression changes- including cancer development, progression, response to treatments, and targeted perturbation-in the interactome network using the concept of cascading failures. The results of these analyses associate robustness with cancer and identify autophagy as an opposite condition. Next, we hypothesized that a comprehensive measure of cancer cell status or function applicable to predicting therapy response should integrate and/or partially reflect the fundamental hallmarks of cancer. To address these questions, we have developed a novel weighted network score, which is used to show that cancer network activity is associated with therapeutic response and synergism., El càncer és una família de malalties complexes que es caracteritzen per una proliferació cel·lular descontrolada i il·limitada i la capacitat de les cèl·lules d'invadir teixit veí i metastatitzar cap a teixit i òrgans llunyans. El desenvolupament i progressió d'un càncer és un procés complex de diversos passos en els quals les cèl·lules pateixen i acumulen mutacions gèniques. Aquestes mutacions alteren els mecanismes cel·lulars fins que la cèl·lula cancerosa ha acumulat les alteracions necessàries per proliferar de manera descontrolada i adquirir la capacitat de colonitzar teixits o òrgans llunyans. Les mutacions que pateixen les cèl·lules durant el procés de la carcinogènesi afecten a diferents nivells del sistema cel·lular. Aquestes mutacions alteren els perfils d'expressió gènica i produeixen canvis quantitatius i qualitatius en el producte gènic (ARN i proteïna) que poden alterar les xarxes complexes cel·lulars per les quals es regulen els mecanismes cel·lulars. En els darrers anys, diverses anàlisis, que han integrat dades d'expressió gènica, han sigut útils per identificar signatures de gens pel pronòstic del càncer o determinar teràpies pel seu tractament. Tanmateix, a causa de la diversitat biològica de la cèl·lula, signatures de gens identificades mitjançant estudis independents sovint tenen pocs o cap gen en comú. Amb la integració de dades d'expressió gènica i d'interaccions entre proteïnes, s'ha pogut explicar la diversitat biològica que capturen aquestes anàlisis mitjançant la identificació de signatures aparentment diferents. Malgrat que la xarxa d'interaccions entre proteïnes (xarxa de l'interactoma) aporta informació addicional del sistema, la relació amb la dinàmica molecular del càncer roman desconeguda. Les cèl·lules canceroses sovint responen a la teràpia generant resistència, ja sigui per la pròpia heterogeneïtat de la cèl·lula cancerosa o adquirint noves mutacions o alteracions en els seus mecanismes. Els canvis adquirits poden afectar a processos biològics o vies de senyalització immerses en les xarxes complexes moleculars ---desregulant-les o modificant la seva activitat--- augmentant la robustesa característica del càncer. En aquest sentit, el desenvolupament d'eines per capturar l'activitat de les xarxes moleculars i per les vies que la cèl·lula adquireix resistència o per les quals pot ser vulnerable, pot ser de gran utilitat per identificar teràpies específiques dirigides a dianes moleculars concretes. En aquest treball, per explorar la relació entre la xarxa de l'interactoma i la dinàmica molecular del càncer, s'ha realitzat una anàlisi toplògica de la xarxa de l'interactoma respecte dels estadis del càncer, pertorbació de gens centrals en el càncer i la resposta al tractament. Per fer-ho, s'ha implementat un model de cascada de fallades que modelitza la propagació de càrrega proteica a través de la xarxa. Els resultats obtinguts mostren que la xarxa de l'interactoma és robusta respecte de les condicions del càncer. Per altra banda, per explorar l'activitat de la xarxa de les cèl·lules canceroses i la seva relació a la sensibilitat de les cèl·lules respecta al tractament, s'ha definit una mesura de l'activitat de la xarxa i s'ha aplicat a les xarxes de diferents càncers. Tot seguit, s'han comparat les mesures obtingudes amb la resposta de les cèl·lules canceroses al tractament de diversos fàrmacs. Els resultats obtinguts, mostren que aquesta mesura diferencia fàrmacs segons les seves característiques, per exemple si són citotòxics o tenen dianes específiques, i que prediu combinacions de fàrmacs amb un efecte sinèrgic.

Lung cancer (LC) remains the leading cause of cancer mortality worldwide, and non-small cell LC (NSCLC) represents 80% of all LC. Oxidative stress and inflammation, autophagy, ubiquitin-proteasome system, nuclear factor (NF)-KB, and mitogen activated protein kinases (MAPK) participate in LC pathophysiology. Currently available treatment for LC is limited and in vivo models are lacking. We hypothesized that antioxidants and NF-KB, MAPK, and proteasome inhibitors may exert an antitumoral response through attenuation of several key biological mechanisms that promote tumorigenesis and cancer cell growth. Body and tumor weights, oxidative stress, antioxidants, inflammation, NF-KB p65 expression, fibulins, apoptosis, autophagy, tumor and stroma histology were evaluated in the subcutaneous tumor of LC (LP07) adenocarcinoma) BALB/c mice, with and without concomitant treatment with NF-KB (sulfasalazine), MEK (U0126), and proteasome (bortezomib) inhibitors, and N-acetyl cysteine (NAC). Compared to LC control mice, in subcutaneous tumors, the four pharmacological agents reduced oxidative stress markers and tumor proliferation (ki-67). Inflammation and NF-KB p65 expression were attenuated by NF-KB and MAPK inhibitors, and the latter also enhanced apoptotic markers. Catalase was induced by the three inhibitors, while bortezomib also promoted superoxide dismutase expression. NF-KB and MEK inhibitors significantly reduced tumor burden through several biological mechanisms that favored tumor degradation and attenuated tumor proliferation. These two pharmacological agents may enhance the anti-tumor activity of selectively targeted therapeutic strategies for LC. Proteasomal inhibition using bortezomib rather promotes tumor degradation, while treatment with antioxidants cannot be recommended. This experimental model supports the use of adjuvant drugs for the improvement of LC treatment. Fil: Mateu-Jimenez, Mercè. Barcelona Biomedical Research Park; España Fil: Fermoselle, Clara. Barcelona Biomedical Research Park; España Fil: Rojo, Federico. Fundación Jiménez Díaz; España Fil: Mateu, Javier. Hospital del Mar; España Fil: Peña, Raúl. Hospital del Mar; España Fil: Urtreger, Alejandro Jorge. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Diament, Miriam. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina Fil: Bal, Elisa Dora. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina Fil: Pijuan, Lara. Hospital del Mar; España Fil: Herreros, Antonio. Hospital del Mar; España Fil: Barreiro, Esther. Barcelona Biomedical Research Park; España

BACKGROUND: Follicular lymphoma (FL) is the second most common type of lymphoid cancer in Western Europe. OBJECTIVE: The aim of this study was to evaluate the cost utility of rituximab-bendamustine treatment compared with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone) treatment as a first-line therapy for patients with advanced FL in Spain. METHODS: A Markov model was developed to estimate the cost effectiveness of rituximab-bendamustine compared with R-CHOP as first-line treatment for patients with advanced FL in the Spanish National Health System (NHS). Transitions between health states (progression-free, including induction and maintenance; first relapse; second relapse; and death) were allowed for the patient cohort in 4-week-long cycles. Clinical data for the extrapolation of progression-free survival curves were obtained from randomized trials. Mortality rates and utilities were obtained from the literature. Outcomes were measured as quality-adjusted life-years (QALYs). The total costs (€, 2013) included drug costs (ex-factory prices with mandatory deductions), disease management costs and adverse event-associated costs. Costs and outcomes were discounted at a 3 % annual rate. Probabilistic sensitivity analysis was performed using 10,000 Monte Carlo simulations to assess the model robustness. RESULTS: Treatment and administration costs during the induction phase were higher for rituximab-bendamustine (€17,671) than for R-CHOP (€11,850). At the end of the 25-year period, the rituximab-bendamustine first-line strategy had a total cost of €68,357 compared with €69,528 for R-CHOP. Health benefits were higher for rituximab-bendamustine treatment (10.31 QALYs) than for R-CHOP treatment (9.82 QALYs). In the probabilistic analysis, rituximab-bendamustine was the dominant strategy over treatment with R-CHOP in 53.4 % of the simulations. CONCLUSION: First-line therapy with rituximab-bendamustine in FL patients was the dominant strategy over treatment with R-CHOP; it showed cost savings and higher health benefits for the Spanish NHS. This work was performed thanks to financial support from Mundipharma Pharmaceuticals, S.L. (Spain). ES and IO are employed by PORIB, a consultant company specialized in health economic evaluations. JMC is employed by Mundipharma Pharmaceuticals. AR has received honoraria from Mundipharma Pharmaceuticals, for advocacy tasks related to the presentwork and investigational funds and lecture honoraria from Roche Pharma. AS and ALG have received honoraria from Mundipharma Pharmaceuticals, for advocacy tasks related to the present work.

Silvia Cufi 1,2 , Bruna Corominas-Faja 1,2 , Alejandro Vazquez-Martin 1,2 , Cristina Oliveras-Ferraros 1,2 , Joan Dorca 2,3 , Joaquim Bosch-Barrera 2,3 , Begona Martin-Castillo 2,4 , Javier A. Menendez 1,2 1 Translational Research Laboratory, Catalan Institute of Oncology (ICO), Girona, Spain 2 Girona Biomedical Research Institute (IDIBGi), Girona, Spain 3 Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain 4 Clinical Research Unit, Catalan Institute of Oncology (ICO), Girona, Spain Received: April 27, 2012, Accepted: April 28, 2012, Published: May 4, 2012, Keywords: Metformin, trastuzumab, HER2, cancer stem cells, breast cancer Correspondence: Javier A. Menendez, email: // // Abstract Trastuzumab-refractory breast cancer stem cells (CSCs) could also explain the high rate of primary resistance to single-agent trastuzumab in HER2 gene-amplified breast cancer patients. The identification of agents with strong selective toxicity for trastuzumab-resistant breast CSCs may have tremendous relevance for how HER2+ breast cancer patients should be treated. Using the human breast cancer cell line JIMT-1, which was established from the pleural metastasis of a patient who was clinically resistant to trastuzumab ab initio , we examined whether preferential killing of the putative CD44 + CD24 -/low breast CSC population might be sufficient to overcome primary resistance to trastuzumab in vivo . Because recent studies have shown that the anti-diabetic biguanide metformin can exert antitumor effects by targeted killing of CSC-like cells, we explored whether metformin’s ability to preferentially kill breast cancer initiating CD44 + CD24 -/low cells may have the potential to sensitize JIMT-1 xenograft mouse models to trastuzumab. Upon isolation for breast cancer initiating CD44 + CD24 -/low cells by employing magnetic activated cell sorting, we observed the kinetics of metformin-induced killing drastically varied among CSC and non-CSC subpopulations. Metformin’s cell killing effect increased dramatically by more than 10-fold in CD44 + CD24 -/low breast CSC cells compared to non-CD44 + CD24 -/low immunophenotypes. While seven-weeks treatment length with trastuzumab likewise failed to reduce tumor growth of JIMT-1 xenografts, systemic treatment with metformin as single agent resulted in a significant two-fold reduction in tumor volume. When trastuzumab was combined with concurrent metformin, tumor volume decreased sharply by more than four-fold. Given that metformin-induced preferential killing of breast cancer initiating CD44 + CD24 -/low subpopulations is sufficient to overcome in vivo primary resistance to trastuzumab, the incorporation of metformin into trastuzumab-based regimens may provide a valuable strategy for treatment of HER2+ breast cancer patients.

Ribonucleases are proteins whose use is promising in anticancer therapy. We have previously constructed different human pancreatic ribonuclease variants that are selectively cytotoxic for tumor cells by introducing a nuclear localization signal into their sequence. However, these modifications produced an important decrease in their stability compromising their behavior in vivo. Here, we show that we can significantly increase the thermal stability of these cytotoxic proteins by introducing additional disulfide bonds by site-directed mutagenesis. One of these variants increases its thermal stability by around 17 &deg, C, without affecting its catalytic activity while maintaining the cytotoxic activity against tumor cells. We also show that the most stable variant is significantly more resistant to proteolysis when incubated with proteinase K or with human sera, suggesting that its half-live could be increased in vivo once administered.

Treball de fi de grau en Biomèdica Tutors: Antoni Ivorra, Quim Castellví In the last years irreversible electroporation techniques (IRE) has started to gain importance exponentially in the medicine field, from being considered as an undesirable side-effect of reversible electroporation treatments to being considered nowadays as a viable treatment against cancer by its right own. IRE treatments are based in electroporation, electrical pulses are delivered to damage the cell membranes and cause the cell death. It has numerous advantages compared with other ablation techniques since is a non-thermal ablation, thus, the tissue scaffold is spared when the treatment is applied. Is possible to predict the extension and shape of the treatment by means of mathematical simulations, the models used to predict that have been experimentally refined. We present an improved method to evaluate in three-dimensions the results of IRE using vegetal models. The technique consists on using a dye solution to enhance the IRE area in sliced potato tubers. After slice digitalization, the electroporated area was automatically identified and the resulting treated volume was calculated. In addition, a threedimensional/nreconstruction of both healthy tissue and IRE volume was generated. The proposed evaluation technique was used to assess different pulse protocols outcomes. Finally, numerical simulations had been carried out to compare the numerical predictions to the experimental observations. The obtained results shows a clearly match between experimental and simulated volumes/nconfirming the reliability of the method proposed.

El tercer càncer més comú avui dia és el càrcer colorectal. Existeixen diverses teràpies per combatre’l com la radioteràpia i la quimioteràpia. L’eficàcia de les teràpies anticancerígenes es pot veure disminuïda o anul·lada per una característica de les cèl·lules cancerígenes, la hipòxia. Ha sorgit recentment una teràpia alternativa per als pacients amb càncer de colon que no pot ser operat. Aquesta utilitza fonts de llum per eliminar el teixit cancerós mitjançant la producció d’oxigen singlet i altres espècies de ERO, s’anomena teràpia fotodinàmica (TF). No obstant, la seva efectivitat també es pot veure afectada per les cèl·lules tumorals hipòxiques. Per millorar la seva eficàcia en el tractament del càncer colorectal s’ha utilitzat la teràpia d’oxigen hiperbàric (TOHB), que elimina l’estat d’hipòxia cel·lular. La combinació de les teràpies ha sigut aplicada utilitzant diverses condicions i el seu efecte s’ha analitzat mitjançant l’assaig MTT i l’anàlisi de cicle cel·lular i de viabilitat cel·lular per citometria. Els resultats obtinguts donen una visió esperançadora de la combinació de TF i TOHB i en anàlisis futurs, s’haurà de corroborar els resultats de la teràpia de TF seguida de TOHB durant 60 minuts.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.-- et al., [Introduction]: Endocrine therapies targeting cell proliferation and survival mediated by estrogen receptor α (ERα) are among the most effective systemic treatments for ERα-positive breast cancer. However, most tumors initially responsive to these therapies acquire resistance through mechanisms that involve ERα transcriptional regulatory plasticity. Herein we identify VAV3 as a critical component in this process. [Methods]: A cell-based chemical compound screen was carried out to identify therapeutic strategies against resistance to endocrine therapy. Binding to ERα was evaluated by molecular docking analyses, an agonist fluoligand assay and short hairpin (sh)RNA-mediated protein depletion. Microarray analyses were performed to identify altered gene expression. Western blot analysis of signaling and proliferation markers, and shRNA-mediated protein depletion in viability and clonogenic assays, were performed to delineate the role of VAV3. Genetic variation in VAV3 was assessed for association with the response to tamoxifen. Immunohistochemical analyses of VAV3 were carried out to determine its association with therapeutic response and different tumor markers. An analysis of gene expression association with drug sensitivity was carried out to identify a potential therapeutic approach based on differential VAV3 expression. [Results]: The compound YC-1 was found to comparatively reduce the viability of cell models of acquired resistance. This effect was probably not due to activation of its canonical target (soluble guanylyl cyclase), but instead was likely a result of binding to ERα. VAV3 was selectively reduced upon exposure to YC-1 or ERα depletion, and, accordingly, VAV3 depletion comparatively reduced the viability of cell models of acquired resistance. In the clinical scenario, germline variation in VAV3 was associated with the response to tamoxifen in Japanese breast cancer patients (rs10494071 combined P value = 8.4 × 10-4). The allele association combined with gene expression analyses indicated that low VAV3 expression predicts better clinical outcome. Conversely, high nuclear VAV3 expression in tumor cells was associated with poorer endocrine therapy response. Based on VAV3 expression levels and the response to erlotinib in cancer cell lines, targeting EGFR signaling may be a promising therapeutic strategy. [Conclusions]: This study proposes VAV3 as a biomarker and a rationale for its use as a signaling target to prevent and/or overcome resistance to endocrine therapy in breast cancer., This work was supported by grants from the Eugenio Rodríguez Pascual Foundation (2012, to MAP), the Government of Catalonia (2009-SGR283, to AV and MAP), the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health (R01 DK015556, to JAK), the Red Cooperative Research Thematic Network on Cancer (RTICC) (12/0036/0002 to XRB and 12/0036/0008 to XRB and MAP) and the Spanish Ministry of Health, Fund for Health Research–Institute of Health Carlos III (11/00951 to AU and 12/01528 to MAP).

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence., Glioblastomas are highly diffuse, malignant tumors that have so far evaded clinical treatment. The strongly invasive behavior of cells in these tumors makes them very resistant to treatment, and for this reason both experimental and theoretical efforts have been directed toward understanding the spatiotemporal pattern of tumor spreading. Although usual models assume a standard diffusion behavior, recent experiments with cell cultures indicate that cells tend to move in directions close to that of glioblastoma invasion, thus indicating that a biased random walk model may be much more appropriate. Here we show analytically that, for realistic parameter values, the speeds predicted by biased dispersal are consistent with experimentally measured data. We also find that models beyond reaction-diffusion-advection equations are necessary to capture this substantial effect of biased dispersal on glioblastoma spread. © IOP Publishing and Deutsche Physikalische Gesellschaft., This work was supported by MEC (FIS-2012-31307 grant, JF), the Generalitat de Catalunya (2009-SGR-374 grant, JF), the Marcelino Bot́ín Foundation (RVS) and the Santa Fe Institute (RVS).

Brain tumor segmentation consists of separating the different tumor tissues (solid or active tumor, edema,and necrosis) from normal brain tissues: gray matter (GM), white matter (WM), and cerebrospinal fluid (CSF). In brain tumor studies, the existence of abnormal tissues may be easily detectable most of the time. However, accurate and reproducible segmentation and characterization of abnormalities are not straightforward. In the past, many researchers in the field of medical imaging and soft computing have made significant survey in the field of brain tumor segmentation. Both semiautomatic and fully automatic methods have been proposed. Clinical acceptance of segmentation techniques has depended on the simplicity of the segmentation, and the degree of user supervision. Interactive or semiautomatic methods are likely to remain dominant in practice for some time, especially in these applications where erroneous interpretations are unacceptable. This article presents an overview of the most relevant brain tumor segmentation methods, conducted after the acquisition of the image. Given the advantages of magnetic resonance imaging over other diagnostic imaging, this survey is focused on MRI brain tumor segmentation. Semiautomatic and fully automatic techniques are emphasized.

The recent identification of HER2 gene amplification as a novel predictor of resistance to the EGFR (HER2)-targeted antibody cetuximab and of response to combination therapies against EGFR and HER2 in wild-type KRAS tumor settings may represent a further step toward personalized medicine for patients with colorectal cancer. Herein, we show that transcriptional upregulation of HER2 expression in the absence of HER2 gene amplification is a molecular phenomenon that takes place in EGFR-dependent, wild-type KRAS squamous cell carcinoma (SCC) cells that acquire resistance to cetuximab. Since cetuximab activity against cetuximab-refractory SCC cells can be fully restored in the presence of the anti-HER2 monoclonal antibody trastuzumab, our findings suggest that, beyond HER2 gene amplification, we might need to redefine the threshold values for HER2 positivity to improve the accuracy of the selection of cetuximab-refractory patients with wild-type KRAS that may benefit from receiving a cetuximab/trastuzumab combination.

Ribonucleases are promising agents for use in anticancer therapy. Engineering a nuclear localization signal into the sequence of the human pancreatic ribonuclease has been revealed as a new strategy to endow this enzyme with cytotoxic activity against tumor cells. We previously described a cytotoxic human pancreatic ribonuclease variant, named PE5, which is able to cleave nuclear RNA, inducing the apoptosis of cancer cells and reducing the amount of P-glycoprotein in different multidrug-resistant cell lines. These results open the opportunity to use this ribonuclease in combination with other chemotherapeutics. In this work, we have investigated how to improve the properties of PE5 as an antitumor drug candidate. When attempting to develop a recombinant protein as a drug, two of the main desirable attributes are minimum immunogenicity and maximum potency. The improvements of PE5 have been designed in both senses. First, in order to reduce the potential immunogenicity of the protein, we have studied which residues mutated on PE5 can be reverted to those of the wild-type human pancreatic ribonuclease sequence without affecting its cytotoxicity. Second, we have investigated the effect of introducing an additional nuclear localization signal at different sites of PE5 in an effort to obtain a more cytotoxic enzyme. We show that the nuclear localization signal location is critical for the cytotoxicity. One of these variants, named NLSPE5, presents about a 10-fold increase in cytotoxicity respective to PE5. This variant induces apoptosis and kills the cells using the same mechanism as PE5 This work has been supported by Grant BFU2009-06935 from MICINN (Spain) and by Grant GRCT04 from the University of Girona. We are very grateful to Dr. Milica Pesic and Dr. Sabera Ruzdijićfor providing us with the NCI-H460/R cell line and to Dr. Ramon Colomer for providing us with the NCI/ ADR-RES cell line. A.V., J.C., and P.T. acknowledge their fellowship from MINEDU, Universitat de Girona, and MEC (Spain), respectively

Ribonucleases are small basic proteins that have shown remarkable antitumor activity linked to their ability to destroy RNA. Therefore, they are a second line of cancer chemotherapeutics as they are not genotoxic. This chapter summarizes the main biochemical characteristics of these enzymes and the key factors responsible for their cytotoxic mechanism. Some of them are shared by most cytotoxins, but each RNase has particular cancer cell killing abilities. The effects on the cell cycle and the induced apoptosis mechanism are cell dependent. The knowledge obtained from the cytotoxic mechanism of natural cytotoxic RNases has been used to artificially engineer more potent and selective RNA-degrading enzymes. These approaches are also described. The chapter ends with a brief description of the results of the clinical trials performed with RNases This work has been supported by grants BFU2009-06935 from MICINN (Spain) and GRCT04 from the University of Girona

Introduction. Inhibiting the enzyme Fatty Acid Synthase (FASN) leads to apoptosis of breast carcinoma cells, and this is linked to human epidermal growth factor receptor 2 (HER2) signaling pathways in models of simultaneous expression of FASN and HER2. Methods. In a xenograft model of breast carcinoma cells that are FASN+ and HER2+, we have characterized the anticancer activity and the toxicity profile of G28UCM, the lead compound of a novel family of synthetic FASN inhibitors. In vitro, we have analyzed the cellular and molecular interactions of combining G28UCM with anti-HER drugs. Finally, we have tested the cytotoxic ability of G28UCM on breast cancer cells resistant to trastuzumab or lapatinib, that we developed in our laboratory. Results. In vivo, G28UCM reduced the size of 5/14 established xenografts. In the responding tumours, we observed inhibition of FASN activity, cleavage of poly- ADPribose polymerase (PARP) and decrease of p-HER2, p- protein kinase B (AKT) and p-ERK1/2, which were not observed in the nonresponding tumours. In the G28UCM-treated animals, no significant toxicities occurred, and weight loss was not observed. In vitro, G28UCM showed marked synergistic interactions with trastuzumab, lapatinib, erlotinib or gefitinib (but not with cetuximab), which correlated with increases in apoptosis and with decreases in the activation of HER2, extracellular signal-regulated kinase (ERK)1/2 and AKT. In trastuzumab-resistant and in lapatinib-resistant breast cancer cells, in which trastuzumab and lapatinib were not effective, G28UCM retained the anticancer activity observed in the parental cells. Conclusions. G28UCM inhibits fatty acid synthase (FASN) activity and the growth of breast carcinoma xenografts in vivo, and is active in cells with acquire resistance to anti-HER2 drugs, which make it a candidate for further pre-clinical development

Projecte final de carrera fet en col.laboració amb Université catholique de Louvain. Ecole Polytechnique de Louvain Breast cancer is one of the major causes of death among women. Nowadays screening mammography is the most adopted technique to perform an early breast cancer detection ahead other procedures like screen film mammography (SFM) or ultrasound scan. Computer assisted diagnosis (CAD) of mammograms attempts to help radiologists providing an automatic procedure to detect possible cancers in mammograms. Suspicious breast cancers appear as white spots in mammograms, indicating small clusters of micro-calcifications. Mammogram sensitivity decreases due some factors like density of the breast, presence of labels, artifacts or even pectoral muscle. The pre-processing of mammogram images is a very important step in the breast cancer analysis and detection because it might reduce the number of false positives. In this thesis we propose a method to segment and classify automatically mammograms according to their density. We perform several procedures including pre-processing (enhancement of the image, noise reduction, orientation finding or borders removal) and segmentation (separate the breast from the background, labels and pectoral muscle present in the mammograms) in order to increase the sensitivity of our CAD system. The final goal is the classification for diagnosis, in other words, finding the density class for an incoming mammography in order tot determine if more tests are needed to find possible cancers in the image. This functionality will be included in a new clinical imaging annotation system for computer aided breast cancer screening developed by the Communications and Remote Sensing Department at the Université Catholique de Louvain. The source code for the pre-processing and segmentation step has been programmed in C++ using the library of image processing ITK and CMake compiler. The performed method has been applied to medio-lateral oblique-view (MLO) mammograms as well as on caniocauldal mammograms (CC) belonging to different databases. The classification step has been implemented in Matlab. We have tested our pre-processing method obtaining a rate of 100% success removing labels and artifacts from mammograms of mini-MIAS database. The pectoral removal rate has been evaluated subjectively obtaining a rate of good removal of 57.76%. Finally, for the classification step, the best recognition rate that we have obtained was 76.25% using only pixel values, and 77.5% adding texture features, classifying images belonging to mini-MIAS database into 3 different density types. These results can be compared with the actual state of the art in segmentation and classification of biomedical images. El cáncer de mama es una de las mayores causas de muerte entre las mujeres. Actualmente, las mamografías digitales son la técnica más adoptada para realizar una previa detección de estos cánceres antes que otros procedimientos como "screen film mammography (SFM)" o escáneres de ultrasonidos. Los programas de diagnóstico automático (CAD) ayudan a los radiólogos proveyéndolos de un procedimiento automático para detectar posibles cánceres en las mamografías. Posibles cánceres aparecen en las mamografías como puntos blancos indicando pequeños grupos de micro-calcificaciones. La sensibilidad de las mamografias decrece debido a algunos factores como la densidad del pecho, presencia de etiquetas o artefactos o incluso de músculo pectoral. El pre-procesado de las mamografías es un paso muy importante en la detección de posibles cánceres de mama ya que puede reducir el número de falsos positivos. En esta tesis proponemos un método para segmentar y clasificar automáticamente las mamografías según su densidad. Hemos realizado varios procedimientos incluyendo, pre-procesado (realce de la imagen, reducción de ruido, descubrimiento de la orientación o supresión de bordes) y segmentación (separar el pecho de fondo, etiquetas y músculo pectoral presente en mamografías) para incrementar la sensibilidad de nuestro sistema CAD. El objetivo final es la clasificación para diagnosis, en otras palabras, encontrar la clase de densidad para una mamografía entrante y determinar si son necesarios más pruebas para encontrar posibles cánceres en las imágenes. Esta funcionalidad va a ser incluida en una nueva aplicación ara anotación de imágenes clínicas desarrollada por el Departamento de Comunicación y Detección Remota de la Universidad Católica de Lovaina. El código fuente para el pre-procesado y la segmentación ha sido desarrollado en C++ utilizando la librería de procesado de imagen ITK y el compilador CMake. El método implementado puede ser aplicado a tanto medio-lateral (MLO) como a caniocauldal mamografías (CC) pertenecientes a diferentes bases de datos. El método de clasificación ha sido implementado en Matlab. Hemos testeado nuestro método de pre-procesado obteniendo una tasa de suceso próxima al 100% en la eliminación de etiquetas y artefactos de la base de datos de mamografías mini-MIAS. La tasa de supresión de músculo pectoral ha sido evaluada de forma subjetiva obteniendo un 57.76%. Finalmente, en el método de clasificación se ha obtenido un 76.25% usando sólo información de los píxeles y un 77.5% usando información de texturas. Los resultados pueden ser comparados con el actual estado del arte en segmentación y clasificación de imágenes biomédicas. El càncer de mama és una de les majors causes de mort entre les dones. Actualment, les mamografies digitals són la tècnica més utilitzada per realitzar una prèvia detecció d'aquests càncers abans que altres procediments com "screen film mammography (SFM)" o escàners d'ultrasons. Els programes de diagnòstic automàtic (CAD) ajuden als radiòlegs proveïnt d'un procediment automàtic per detectar possibles càncers a les mamografies. Possibles càncers apareixen en les mamografies com punts blancs indicant petits grups de micro-calcificacions. La sensibilitat de les mamografies decreix a causa d'alguns factors com la densitat del pit, presència d'etiquetes o artefactes o fins i tot de múscul pectoral. El pre-processat de les mamografies és un pas molt important en la detecció de possibles càncers de mama ja que pot reduir el nombre de falsos positius. En aquesta tesi proposem un mètode per segmentar i classificar automàticament les mamografies segons la seva densitat. Hem realitzat diversos procediments incloent, pre-processat (realç de la imatge, reducció de soroll, descobriment de l'orientació o supressió de vores) i segmentació (separar el pit de fons, etiquetes i múscul pectoral present en mamografies) per incrementar la sensibilitat de nostre sistema CAD. L'objectiu final és la classificació per diagnosi, en altres paraules, trobar la classe de densitat per a una mamografia entrant i determinar si són necessaris més proves per trobar possibles càncers en les imatges. Aquesta funcionalitat serà inclosa en una nova aplicació ara anotació d'imatges clíniques desenvolupada pel Departament de Comunicació i Detecció Remota de la Universitat Catòlica de Lovaina. El codi font per al pre-processat i la segmentació ha estat desenvolupat en C + + utilitzant la llibreria de processat d'imatge ITK i el compilador CMake. El mètode implementat pot ser aplicat a tant mediolateral (MLO) com a caniocauldal mamografies (CC) pertanyents a diferents bases de dades. El mètode de classificació ha estat implementat en Matlab. Hem testejat el nostre mètode de pre-processat obtenint una taxa de succés propera al 100% en l'eliminació d'etiquetes i artefactes de la base de dades de mamografies mini-MIAS. La taxa de supressió de múscul pectoral ha estat avaluada de manera subjectiva obtenint un 57.76%. Finalment, en el mètode de classificació s'ha obtingut un 76.25% usant només informació dels píxels i un 77.5% usant informació de textures. Els resultats poden ser comparats amb l'actual estat de l'art en segmentació i classificació d'imatges biomèdiques.

Includes supplementary materials for the online appendix. Includes supplementary materials for the online appendix. Anaplastic lymphoma kinase (ALK) genomic alterations have emerged as a potent predictor of benefit from treatment with ALK inhibitors in several cancers. Currently, there is no information about ALK gene alterations in urothelial carcinoma (UC) and its correlation with clinical or pathologic features and outcome. Samples from patients with advanced UC and correlative clinical data were collected. Genomic imbalances were investigated by array comparative genomic hybridization (aCGH). ALK gene status was evaluated by fluorescence in situ hybridization (FISH). ALK expression was assessed by immunohistochemistry (IHC) and high-throughput mutation analysis with Oncomap 3 platform. Next generation sequencing was performed using Illumina Genome Analyzer IIx, and Illumina HiSeq 2000 in the FISH positive case. 70 of 96 patients had tissue available for all the tests performed. Arm level copy number gains at chromosome 2 were identified in 17 (24%) patients. Minor copy number alterations (CNAs) in the proximity of ALK locus were found in 3 patients by aCGH. By FISH analysis, one of these samples had a deletion of the 5′ALK. Whole genome next generation sequencing was inconclusive to confirm the deletion at the level of the ALK gene at the coverage level used. We did not observe an association between ALK CNA and overall survival, ECOG PS, or development of visceral disease. ALK genomic alterations are rare and probably without prognostic implications in UC. The potential for testing ALK inhibitors in UC merits further investigation but might be restricted to the identification of an enriched population.

Ribonucleases represent a new class of antitumor RNA-damaging drugs. However, many wild-type members of the vertebrate secreted ribonuclease family are not cytotoxic because they are not able to evade the cytosolic ribonuclease inhibitor. We previously engineered the human pancreatic ribonuclease to direct it to the cell nucleus where the inhibitor is not present. The best characterized variant is PE5 that kills cancer cells through apoptosis mediated by the p21WAF1/CIP1 induction and the inactivation of JNK. Here, we have used microarray-derived transcriptional profiling to identify PE5 regulated genes on the NCI/ADR-RES ovarian cancer cell line. RT-qPCR analyses have confirmed the expression microarray findings. The results show that PE5 cause pleiotropic effects. Among them, it is remarkable the down-regulation of multiple genes that code for enzymes involved in deregulated metabolic pathways in cancer cells This work has been supported by grants BFU2009-06935 and BIO2013-43517 from MINECO (Spain) and SING12/0 from UdG (Spain).

Cell 166(3), 740-754 (2016). doi:10.1016/j.cell.2016.06.017, Published by Elsevier, New York, NY

// Elisabet Cuyas 1,2 , Salvador Fernandez-Arroyo 3 , Bruna Corominas-Faja 1,2 , Esther Rodriguez-Gallego 3 , Joaquim Bosch-Barrera 2,4 , Begona Martin-Castillo 2,5 , Rafael De Llorens 6 , Jorge Joven 3 and Javier A. Menendez 1,2 1 Metabolism and Cancer Group, Translational Research Laboratory, Catalan Institute of Oncology (ICO), Girona, Catalonia, Spain 2 Molecular Oncology Group, Girona Biomedical Research Institute (IDIBGI), Girona, Catalonia, Spain 3 Unitat de Recerca Biomedica (URB-CRB), Institut d’Investigacio Sanitaria Pere i Virgili (IISPV), Universitat Rovira i Virgili, Reus, Catalonia. Spain 4 Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Catalonia, Spain 5 Clinical Research Unit, Catalan Institute of Oncology (ICO), Girona, Catalonia, Spain 6 Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Girona, Catalonia, Spain Correspondence to: Javier A. Menendez, email: // Keywords : IDH1, oncometabolites, metformin, anaplerosis, cancer Received : January 17, 2015 Accepted : March 11, 2015 Published : March 30, 2015 Abstract Metabolic flexibility might be particularly constrained in tumors bearing mutations in isocitrate dehydrogenase 1 (IDH1) leading to the production of the oncometabolite 2-hydroxygluratate (2HG). To test the hypothesis that IDH1 mutations could generate metabolic vulnerabilities for therapeutic intervention, we utilized an MCF10A cell line engineered with an arginine-to-histidine conversion at position 132 (R132H) in the catalytic site of IDH1, which equips the enzyme with a neomorphic α-ketoglutarate to 2HG reducing activity in an otherwise isogenic background. IDH1 R132H/+ and isogenic IDH1 +/+ parental cells were screened for their ability to generate energy-rich NADH when cultured in a standardized high-throughput Phenotype MicroArrayplatform comprising >300 nutrients. A radical remodeling of the metabotype occurred in cells carrying the R132H mutation since they presented a markedly altered ability to utilize numerous carbon catabolic fuels. A mitochondria toxicity-screening modality confirmed a severe inability of IDH1-mutated cells to use various carbon substrates that are fed into the electron transport chain at different points. The mitochondrial biguanide poisons, metformin and phenformin, further impaired the intrinsic weakness of IDH1-mutant cells to use certain carbon-energy sources. Additionally, metabolic reprogramming of IDH1-mutant cells increased their sensitivity to metformin in assays of cell proliferation, clonogenic potential, and mammosphere formation. Targeted metabolomics studies revealed that the ability of metformin to interfere with the anaplerotic entry of glutamine into the tricarboxylic acid cycle could explain the hypersensitivity of IDH1-mutant cells to biguanides. Moreover, synergistic interactions occurred when metformin treatment was combined with the selective R132H-IDH1 inhibitor AGI-5198. Together, these results suggest that therapy involving the simultaneous targeting of metabolic vulnerabilities with metformin, and 2HG overproduction with mutant-selective inhibitors (AGI-5198-related AG-120 [Agios]), might represent a worthwhile avenue of exploration in the treatment of IDH1-mutated tumors.

Pregunta d’investigació: És més efectiva la combinació d’exercici terapèutic i Mindfulness envers únicament activitat física per a l’abordatge simptomàtic en pacients amb càncer de mama tractats amb quimioteràpia?. Objectius: Com a objectiu general, valorar la efectivitat d’un tractament basat en la combinació d’exercici terapèutic combinat amb Mindfulness per l’abordatge simptomàtic en pacients amb càncer de mama tractats amb quimioteràpia. I com a objectiu específic valorar la fatiga, qualitat del son, el dolor i la depressió en aquests pacients. Metodologia: es realitzarà un estudi controlat aleatoritzat amb una mostra de 64 pacients en període de reminiscència de càncer de mama els quals dividirem en dos grups de 32 persones. La intervenció té una durada de 8 setmanes. El grup control realitzarà activitat terapèutica ( 3 dies per setmana 1h cada sessió) i el grup experimental realitzarà activitat terapèutica combinada amb treball formal Mindfulness (90 minuts setmanalment, un total de 12 hores) i treball informal de 45 minuts diaris. Es valorarà la fatiga mitjançant el qüestionari de Piper de la Fatiga, la qualitat del son mitjançant l’índex de la qualitat del son de Pittsburgh, el dolor mitjançant l’escala EVA i la depressió mitjançant l’escala de Hamilton per a la Depressió HAM-D. Es realitzaran 3 valoracions: una pre-tractament, una altra post-tractament a les 8 setmanes d’intervenció i una de llarg termini a l’any des de la primera intervenció. Research question: Is the combination of therapeutic exercise and Mindfulness more effective only for physical activity for the symptomatic approach in breast cancer patients treated with chemotherapy? Objectives: As a general objective, assess the effectiveness of a treatment based on the combination of therapeutic exercise combined with Mindfulness for the symptomatic approach in patients with breast cancer treated with chemotherapy. And as a specific objective, assess fatigue, sleep quality, pain and depression in these patients. Methodology: A randomized controlled study will be carried out with a sample of 64 patients reminiscent of breast cancer, which will be divided into two groups of 32 people. The intervention lasts for 8 weeks. The control group will perform therapeutic activity (3 days per week 1h each session) and the experimental group will perform therapeutic activity combined with formal Mindfulness work (90 minutes per week, a total of 12 hours) and informal work 45 minutes daily. Fatigue will be assessed through the Piper Fatigue questionnaire, sleep quality through the Pittsburgh sleep quality index, pain through the EVA scale and depression through the Hamilton scale for depression HAM-D. Three assessments will be made: one pre-treatment, another post-treatment at 8 weeks of intervention and one long-term per year from the first intervention.

Conjugates of a dicarba analogue of octreotide, a potent somatostatin agonist whose receptors are overexpressed on tumor cells, with [PtCl 2(dap)] (dap = 1-(carboxylic acid)-1,2-diaminoethane) (3), [(η 6-bip)Os(4-CO 2-pico)Cl] (bip = biphenyl, pico = picolinate) (4), [(η 6-p-cym)RuCl(dap)] + (p-cym = p-cymene) (5), and [(η 6-p-cym)RuCl(imidazole-CO 2H)(PPh 3)] + (6), were synthesized by using a solid-phase approach. Conjugates 3-5 readily underwent hydrolysis and DNA binding, whereas conjugate 6 was inert to ligand substitution. NMR spectroscopy and molecular dynamics calculations showed that conjugate formation does not perturb the overall peptide structure. Only 6 exhibited antiproliferative activity in human tumor cells (IC 50 = 63 ± 2 μ in MCF-7 cells and IC 50 = 26 ± 3 μ in DU-145 cells) with active participation of somatostatin receptors in cellular uptake. Similar cytotoxic activity was found in a normal cell line (IC 50 = 45 ± 2.6 μ in CHO cells), which can be attributed to a similar level of expression of somatostatin subtype-2 receptor. These studies provide new insights into the effect of receptor-binding peptide conjugation on the activity of metal-based anticancer drugs, and demonstrate the potential of such hybrid compounds to target tumor cells specifically.

The members of the epidermal growth factor (EGF)/ErbB family are prime targets for cancer therapy. However, the therapeutic efficiency of the existing anti-ErbB agents is limited. Thus, identifying new molecules that inactivate the ErbB receptors through novel strategies is an important goal on cancer research. In this study we have developed a shorter form of human EGF (EGFt) with a truncated C-terminal as a novel EGFR inhibitor. EGFt was designed based on the superimposition of the three-dimensional structures of EGF and the Potato Carboxypeptidase Inhibitor (PCI), an EGFR blocker previously described by our group. The peptide was produced in E. coli with a high yield of the correctly folded peptide. EGFt showed specificity and high affinity for EGFR but induced poor EGFR homodimerization and phosphorylation. Interestingly, EGFt promoted EGFR internalization and translocation to the cell nucleus although it did not stimulate the cell growth. In addition, EGFt competed with EGFR native ligands, inhibiting the proliferation of cancer cells. These data indicate that EGFt may be a potential EGFR blocker for cancer therapy. In addition, the lack of EGFR-mediated growth-stimulatory activity makes EGFt an excellent delivery agent to target toxins to tumours over-expressing EGFR.

Projecte realitzat en col.laboració amb el centre ICFO-Institut de Ciències Fotòniques Emodin (6-methyl-1,3,8 trihydroxyanthraquinone) is considered as a potential candidate for photodynamic therapy for the treatment of cancer. A success in using of the molecule depends among other factors on the knowledge how fast it penetrates through the cell membrane and then diffuses inside the cell. A spatial distribution of emodin inside cancerous cells is also an important factor. The aim of this study is to investigate the diffusion of emodin in cancerous cells by means of a non invasive and sensitive technique: Surface Enhanced Raman scattering which permits us to study the evolution of the small amount of emodin inside single living cells. The experimental results were compared with a theoretical model that simulated the Brownian diffusion of particles. From this comparison and comparing with previous results in fluorescence spectroscopy we have found that: for a long time, emodin is accumulated in the cytoplasm in a region close to the membrane. The result is discussed in terms of emodin being stored in cell organelles located in the cytoplasm.

Purpose: Triple-negative breast cancer (TNBC) lacks an approved targeted therapy. Despite initial good response to chemotherapy, 30% of the patients relapse within 5 years after treatment. EGFR overexpression is a common marker in TNBC, and its expression has been correlated with poor outcome. Inhibition of fatty acid synthase (FASN) activity leads to apoptosis of human carcinoma cells overexpressing FASN. We tested the hypothesis that blocking FASN in combination with anti-EGFR signaling agents would be an effective antitumor strategy in sensitive and chemoresistant TNBC. Experimental Design: Several TNBC cell lines and 29 primary tumors were included to determine whether FASN is a potential target in TNBC. Doxorubicin-resistant TNBC cell lines (231DXR and HCCDXR) have been developed and characterized in our laboratory. Cellular and molecular interactions of anti-FASN compounds (EGCG and C75) with cetuximab were analyzed. In vivo tumor growth inhibition was evaluated after cetuximab, EGCG, or the combination in TNBC orthoxenograft models. Results: TNBC cell lines showed overexpression of FASN enzyme and its inhibition correlated to FASN levels. FASN staining was observed in all of the 29 TNBC tumor samples. In vitro, EGCG and C75 plus cetuximab showed strong synergism in sensitive and chemoresistant cells. In vivo, the combination of EGCG with cetuximab displayed strong antitumor activity against the sensitive and chemoresistant TNBC orthoxenografts, without signs of toxicity. Conclusions: Our results show that the simultaneous blockade of FASN and EGFR is effective in preclinical models of sensitive and chemoresistant TNBC. Clin Cancer Res; 22(18); 4687–97. ©2016 AACR.

Curs 2014-2015, Els avenços en l’estudi de les bases moleculars del càncer han permès el desenvolupament de nous fàrmacs personalitzats a les necessitats de cada pacient, per tal de maximitzar el seu benefici. Estudis recents suggereixen que els exosomes, vesícules que presenten una mida compresa entre els 30 i els 100 nm, secretats per les cèl·lules tumorals i que contenen diferents biomolècules (mARN, miARN, proteïnes oncogèniques i angiogèniques...), poden ser utilitzades per la caracterització biomolecular del càncer i així, poder dissenyar una teràpia apropiada. No obstant això, la manca de sistemes de detecció de petites concentracions d’exosomes en mostres reals és un obstacle a l’hora de desenvolupar un test de diagnòstic, ja que no és possible normalitzar la quantitat de biomarcador respecte al nombre d’exosomes. L’objectiu del projecte en el qual s’engloba aquest Treball de Fi de Grau és el desenvolupament d’un biosensor ràpid, precís i de simple maneig basat en les guies bimodals per a la detecció d’exosomes, mitjançant un immunoassaig dirigit contra proteïnes de membrana, i la posterior detecció de mARN, corresponent a la variant 1 i 3a/b del gen de fusió EML4-ALK en càncer de pulmó, per mitjà d’un assaig genòmic. El biosensor de guies bimodals possibilita la detecció sense cap tipus de marcatge i la seva integració en dispositius point-of-care, on l’anàlisi es pot realitzar sense cap instrumentació addicional. A més a més, la seva elevada sensibilitat podria permetre detectar ARN directament en mostres reals sense cap amplificació. En conclusió, s’ha desenvolupat un biosensor específic per a la detecció d’exosomes amb la capacitat de detectar-los en mostres de medis tumorals amb un límit de detecció de exosomes/mL. A més a més, en aquest treball s’ha aconseguit dissenyar els bioreceptors que possibilitaran la detecció del mARN contingut, fent els primers passos cap al desenvolupament d’un dispositiu integrat que permetrà la normalització del biomarcador respecte al nombre d’exosomes a temps real., Advances in the study of the molecular basis in cancer have enabled the development of new personalized drugs customized for each patient in order to maximize their benefits. Recent studies suggest that exosomes, vesicles which have a size between 30 and 100 nm, secreted by tumor cells and which contain different biomolecules (mRNA, miRNA, oncogenic and angiogenic proteins…),can be used for biomolecular cancer characterization and thus, designing an appropriate therapy. However, the lack in the detection systems of small concentrations of exosomes on real samples is an obstacle in developing a diagnostic test, because it is not possible to standardize the amount of biomarker compared the number of exosomes. The aim of the project included in this Final Year Project is the development of a fast, specific and simple handling biosensor, based on bimodal waveguides biosensors for the detection of exosomes using an immunoassay against membrane proteins, and subsequent detection of mRNA corresponding to variant 1 and 3a/b of the fusion gene EML4-ALK in lung cancer, using a genomic test. Bimodal waveguide biosensor makes possible the detection without labelling and their integration in point-of-care devices, where the analysis can be performed without extra instruments. In addition, its high sensitivity could allow directly detection of RNA in real samples without amplification. In conclusion, we have developed a specific biosensor for exosomes detection with the ability to detect them in tumor cell culture with a limit detection of exosomes/mL. Moreover, in this project we have achieved to design bioreceptors that will enable the detection of mRNA content, giving the first steps for the development of an integrated device that will allow the normalization of the biomarker compared to the number of exosomes at real time.