Search

El càncer és una de les principals causes de mort segons l'OMS, sent el càncer de pulmó el segon més comú a Espanya. El tractament en la major part dels casos implica radioteràpia externa amb feixos de fotons. En el cas del càncer de pulmó, el tumor es troba als pulmons, els quals estan protegits per la caixa toràcica. Així doncs, els feixos de radiació externs han de travessar zones de extrema inhomogeneïtat (os-pulmó). En aquest treball es planteja la comparació de dos algoritmes de planificació comercials per a radioteràpia externa en càncers de pulmó: AAA (el més utilitzat comunament) i Acuros XB (algoritme de major precisió) tots dos d'Eclipse-Varian en maniquins. El codi PRIMO, un sistema de validació per simulació Monte Carlo co-desenvolupat per investigadors de l'INTE-UPC, es prendrà com a referència per seleccionar la planificació més adequada. El cáncer es una de las principales causas de muerte según la OMS, siendo el cáncer de pulmón el segundo más común en España. El tratamiento en la mayor parte de los casos implica radioterapia externa con haces de fotones. En el caso del cáncer de pulmón, el tumor se encuentra en los pulmones, los cuales a su vez están protegidos por la caja torácica. Así, los haces de radiación externos deben atravesar zonas de extrema inhomogeneidad (hueso-pulmón). En este trabajo se plantea la comparación de dos algoritmos de planificación comerciales para radioterapia externa en cánceres de pulmón: AAA (el más utilizado comúnmente) y Acuros XB (algoritmo de mayor precisión) ambos de Eclipse-Varian en maniquís. El código PRIMO, un sistema de validación por simulación Monte Carlo co-desarrollado por investigadores del INTE-UPC, se tomará como referencia para seleccionar la planificación más adecuada. Cancer is one of the leading causes of death according to the WHO, being lung cancer the second most common in Spain. Treatment in most cases involves external radiation therapy with photon beams. In the case of lung cancer, the tumor is found in the lungs, which are protected by the rib cage. Thus, external radiation beams must pass through areas of extreme inhomogeneity (bone-lung). In this work, the comparison of two commercial planning algorithms for external radiotherapy in lung cancers is proposed: AAA (the most commonly used) and Acuros XB (greater precision algorithm) both of Eclipse-Varian in phantoms. The PRIMO code, a Monte Carlo simulation validation system co-developed by INTE-UPC researchers, will be taken as a reference for in order to select the most appropriate planification system.

Apoptosis is a key cell death pathway in mammalian cells. Understanding this process and its regulation has been a subject of study in the last three decades. Members of the Bcl-2 family of proteins are involved in the regulation of apoptosis through mitochondrial poration with the subsequent initiation of apoptosis. Deregulation of proapoptotic proteins contributes to the progression of many tumor processes. Understanding how these pore-forming Bcl-2 proteins Bak and Bax are activated is key to find new anticancer treatments. As no drug capable of activating Bak has been disclosed yet, the study of the structural features of BH3 peptides-known as Bak activators-relevant for binding along with its binding energy decomposition analysis, becomes essential for designing novel small-molecule mimics of BH3. Interestingly, a BH3 Bim analogue-inactivating Bak has recently been discovered, opening a question on the molecular features that determine the functions of BH3 peptides. Therefore, the present work is aimed at understanding the way BH3 peptides activate or inactivate Bak in order to identify differential structural features that can be used in drug design. For this purpose, complexes of Bak with an activator and an inhibitor have been subjected to a molecular dynamics study. Structural differences were assessed by means of the fluctuations of the corresponding principal components. Moreover, the MMPB/GBSA approach was used to compute the binding free energy of the diverse complexes to identify those residues of the BH3 peptide that exhibit the larger contributions to complex formation. The results obtained in this work show differences between activators and inhibitors, both in structural and energetic terms, which can be used in the design of new molecules that can activate or inactivate proapoptotic Bak

En las últimas décadas, la tecnología del plasma no térmico se ha estudiado ampliamente para muchas aplicaciones, entre las cuales, la más prometedora se encuentra en el campo de la medicina. Esta tecnología permite trabajar a temperatura ambiente, con lo cual podemos desarrollar aplicaciones médicas en las que el plasma se aplique a los tejidos humanos. Entre las distintas aplicaciones terapéuticas que se están desarrollando, encontramos el uso del plasma en terapias contra cáncer. Los plasmas no térmicos pueden generar especies reactivas de oxígeno y nitrógeno que tienen la capacidad de interactuar con los sistemas biológicos. En particular, estas especies reactivas permiten matar células cancerosas de manera selectiva sin dañar las células sanas presentes en los tejidos circundantes. El tratamiento de tumores internos más inaccesibles requiere utilizar tratamientos indirectos, en los que se utilizan líquidos tratados con plasma para poder administrar las especies reactivas cerca del tumor. Sin embargo, la inyección de estos líquidos en el cuerpo humano para administrar especies reactivas no es eficiente, ya que las especies reactivas contenidas en ellos son eliminadas rápidamente por los fluidos corporales. El uso de un sistema de administración adecuado donde se pueda generar y almacenar las especies reactivas, podría ofrecer una forma prometedora y viable para tratar el cáncer. En este sentido, los hidrogeles son excelentes candidatos para desarrollar estos sistemas de administración de especies reactivas ya que estos, puede implantarse mediante cirugía o inyectarse antes del proceso de reticulación, para liberar localmente, y de manera controlada, las especies reactivas para que interactúen eficazmente con los tumores. En este contexto, en el presente proyecto se estudiaron soluciones de heparina, condroitina y albúmina como posibles medios para la generación y almacenamiento de especies reactivas derivades del plasma. Para ello, tres soluciones con concentraciones diferentes de cada biopolímero (1 %, 5 %, 10 % para heparina y condroitina y 0.5 %, 1 %, 2.5 % para albúmina) fueron tratadas con plasma para evaluar la influencia del biopolímero en la producción de especies reactivas y así poder seleccionar la concentración óptima para cada biopolímero. Después, se produjeron hidrogeles mezclando los biopolímeros con agarosa al 4 %. Las propiedades mecánicas y la microestructura de los distintos hidrogeles conteniendo heparina, condroitina y albúmina se estudiaron mediante reología y microscopía electrónica de barrido, respectivamente. Finalmente, se evaluaron las propiedades de liberación de las especies reactivas de hidrogeles formados con las soluciones de biopolímeros tratados con plasma en condiciones fisiológicas en una solución salina tamponada con fosfato a 37 °C. En les darreres dècades, la tecnologia del plasma no tèrmic s'ha estudiat àmpliament per a moltes aplicacions, entre les quals la més prometedora es troba en el camp de la medicina. Aquesta tecnologia permet treballar a temperatura ambient, d’aquesta manera podem desenvolupar aplicacions mèdiques en què el plasma s'apliqui als teixits humans. Entre les diferents aplicacions terapèutiques que actualment s'estan desenvolupant trobem l'ús del plasma en teràpies contra càncer. Els plasmes no tèrmics poden generar espècies reactives d’oxigen i nitrogen que tenen la capacitat d’interactuar amb sistemes biològics. En particular, aquestes espècies reactives permeten matar cèl·lules canceroses de manera selectiva sense danyar les cèl·lules sanes presents als teixits circumdants. El tractament de tumors interns més inaccessibles requereix utilitzar tractaments indirectes, en què s'utilitzen líquids tractats amb plasma per poder administrar les espècies reactives a prop del tumor. No obstant això, la injecció d'aquests líquids al cos humà per administrar espècies reactives no és eficient, ja que les espècies reactives contingudes en ells són eliminades ràpidament pels fluids corporals. El desenvolupament d'un sistema d'administració adequat on es puguin generar i emmagatzemar les espècies reactives podria oferir una forma prometedora i viable per tractar el càncer. En aquest sentit, els hidrogels són excel·lents candidats per desenvolupar aquests sistemes d'administració d'espècies reactives ja que aquests es poden implantar mitjançant cirurgia o injectarse abans del procés de reticulació, per alliberar localment i de manera controlada, les espècies reactives perquè interactuïn eficaçment amb els tumors. En aquest context, en el present projecte es van estudiar solucions d'heparina, condroitina i albúmina com a possibles mitjans per generar i emmagatzemar espècies reactives derivades del plasma. Per aquest motiu, tres solucions amb concentracions diferents de cada biopolímer (1 %, 5 %, 10 % per a heparina i condroitina i 0.5 %, 1 %, 2.5 % per a albúmina) van ser tractades amb plasma per avaluar la influència del biopolímer en la producció d'espècies reactives i així poder seleccionar la concentració òptima per a cada biopolímer. Després, es van produir hidrogels barrejant els biopolímers amb agarosa al 4 %. Les propietats mecàniques i la microestructura dels diferents hidrogels contenint heparina, condroitin i albúmina es van estudiar mitjançant reologia i microscopia electrònica d'escombrat, respectivament. Finalment, es van avaluar les propietats d’alliberament de les espècies reactives d’hidrogels formats amb les solucions de biopolímers tractats amb plasma en condicions fisiològiques en una solució salina tamponada amb fosfat a 37 °C. Non-thermal plasma technology has been largely studied in the last decades for many applications, and medicine is among the most promising ones. Working at room temperature allows to treat human tissues and to develop medical applications among them the cancer therapy. Non-thermal plasmas can generate reactive oxygen and nitrogen species that have the ability to interact with biological systems. In particular, they are selective in killing cancer cells without damaging the healthy cells present in the surrounding tissues. Internal cancers require to use indirect treatments and plasma treated liquids. However, the injection of these liquids directly in human body would not be beneficial since the reactive species contained in them would be quickly washed away by the body fluids. Generation and storage of reactive species in suitable delivery systems like hydrogels, could offer a promising and viable way to treat cancers. Indeed, the hydrogels can be implanted by surgery or be injected before crosslinking and release locally the reactive species which can interact effectively with tumours. In this project, solutions of heparin, chondroitin and albumin were studied as possible media for reactive species generation by plasma treatment and storage. Three concentrations of each polymer (1%w/w, 5%w/w, 10%w/w for heparin and chondroitin and 0.5%w/w, 1%w/w, 2.5%w/w for albumin) were plasma treated to assess the influence of polymers on the production of reactive species and to select the optimal concentration for each polymer. Hydrogels were produced by mixing polymers with agarose 4% w/w. The mechanical properties and the microstructure of the hydrogels were studied by rheology and scanning electron microscopy, respectively. Finally, the release properties of hydrogels loaded with plasma-generated reactive species were studied in phosphate buffer saline at 37 °C. Incoming

This project was undertaken in attempt to improve the efficacy of the drug delivery while avoiding systemic toxicity, designing a novel implantable drug delivery platform for iontophoretic brain cancer therapy. 3D fabrication is fast becoming key instrument in the medical field. Thus, we explored microfabrication of an implantable drug delivery device, concretely using direct laser writing to electrostatically deliver charged cancer drugs into brain tumours, technology which is called iontophoresis – basically relies on the motion of ions in electric fields. This technology avoids pressure-related problems, such as drug reflux and brain oedema formation, encountered with convection enhanced delivery by solely relying on the motion of ions in electric fields. The reason for the interest in iontophoresis is that this technique is very versatile allowing to deliver substances into tissue which are usually impermeable. Additionally, it opens up the possibility to deliver drugs in a targeted fashion directly into the desired tissue instead of administering it systemically which reduces off-target toxicity and allows higher delivery rates than when relying solely on passive diffusion of the drug. Overall, this study strengthens the idea that very precise microdevices can be fabricated using Nanoscribe while meeting the biocompatibility criteria, which opens the door to future applications in other areas such as neurological diseases and other cancer treatments Aquest projecte es va dur a terme per intentar millorar l'eficàcia del lliurament de fàrmacs, evitant la toxicitat sistèmica, dissenyant una nova plataforma de lliurament de fàrmacs implantables en malalts de càncer cerebral. Concretament es va utilitzar escriptura làser directa per lliurar electrostàticament fàrmacs contra el càncer amb càrrega als tumors cerebrals, tecnologia que s'anomena iontoforesi: bàsicament es basa en el moviment dels ions en camps elèctrics. Aquesta tecnologia evita problemes relacionats amb la pressió, com ara el reflux de fàrmacs i la formació d'edema cerebral. La raó per l'interès de la iontoforesi és que aquesta tècnica és molt versàtil i permet transferir substàncies al teixit que solen ser impermeables. A més, obre la possibilitat de lliurar medicaments de manera específica directament al teixit desitjat en lloc d'administrar-lo d’una forma sistèmica, fet que redueix la toxicitat fora de l'objectiu i permet taxes de lliurament més altes que quan es confia només en la difusió passiva del fàrmac. En general, aquest estudi reforça la idea que es poden fabricar microdispositius molt precisos que compleixen els criteris de biocompatibilitat. Aquest fet obre la porta al futures aplicacions en altres àrees com les malalties neurològiques i altres tractaments contra el càncer

Different EGFR tyrosine kinase inhibitors (TKIs) have been developed for the treatment of non-small cell lung cancer (NSCLC) patients harboring sensitizing mutations in the EGFR gene. Apart from acquired secondary mutations, multiple resistance mechanisms have been reported, such as the overexpression of fatty acid synthase (FASN), a multi-functional enzyme essential for the de novo lipogenesis, or the increase of cancer stem cells, a small subpopulation within the tumor responsible for relapse, metastasis, and resistance to therapies. Hence, the purpose of this work is to evaluate the novel FASN inhibitor AZ12756122, both alone and in combination with gefitinib and osimertinib, in EGFR-mutated (EGFRm) lung adenocarcinoma cell models sensitive and resistant to EGFR-TKIs. The molecular effect of AZ12756122 (alone and in combination with EGFR-TKI) on FASN, EGFR/ STAT3, Akt/mTOR, and MAPK signaling pathways was analyzed using RT-qPCR and Western blot. FASN expression was also evaluated in samples from patients with EGFRm NSCLC through immunohistochemistry. Our findings revealed that AZ12756122 caused cytotoxic effects inducing apoptosis, downregulated FASN expression and activity, decreased the activation of EGFR and Akt/mTOR pathway, and reduced cancer stem-like cells. Furthermore, the combination of AZ12756122 and osimertinib sensitized cells to EGFR-TKI, showing a syner- gistic effect that resulted in a reduction in the activation of EGFR, Akt/mTOR, and MAPK signaling pathways. Our study also showed that FASN+ EGFRm NSCLC patients exhibited a longer mPFS in patients who responded to EGFR-TKI treatment. In conclusion, FASN inhibition should be further studied for the treatment, alone or in combination with EGFR-TKIs, for EGFRm NSCLC patients

Simple Summary During the past decades the achievements made in treating cancers have significantly improved the survival of patients. However, cancer is still one of the leading causes of mortality. It is suggested that treatment failure is mediated by a subpopulation of tumor cells named cancer stem cells that can survive after treatment and promote cancer relapse. Targeting these cells is important to improve cancer therapy. The aim of our study is to determine the effect of a human ribonuclease variant on breast cancer cells grown in 3D and on cancer stem cells. Moreover, we study its effect on the ability of breast cancer cells to migrate and produce metastasis, responsible for about 90% of cancer deaths. We show that this ribonuclease arrests tumor cells grown in 3D without affecting normal breast cells, and this significantly inhibits cancer stem cell development. Additionally, it reduces the migratory and invasive capacities of tumor cells. Abstract Despite the significant advances in cancer research made in recent years, this disease remains one of the leading causes of death worldwide. In part, this is due to the fact that after therapy, a subpopulation of self-renewing tumor cells can survive and promote cancer relapse, resistance to therapies and metastasis. Targeting these cancer stem cells (CSCs) is therefore essential to improve the clinical outcome of cancer patients. In this sense, multi-targeted drugs may be promising agents targeting CSC-associated multifocal effects. We have previously constructed different human pancreatic ribonuclease (RNase) variants that are cytotoxic for tumor cells due to a non-classical nuclear localization signal introduced in their sequence. These cytotoxic RNases affect the expression of multiple genes involved in deregulated metabolic and signaling pathways in cancer cells and are highly cytotoxic for multidrug-resistant tumor cell lines. Here, we show that these cytotoxic nuclear-directed RNases are highly selective for tumor cell lines grown in 3D, inhibit CSCs’ development and diminish the self-renewal capacity of the CSCs population. Moreover, these human RNase variants reduce the migration and invasiveness of highly invasive breast cancer cells and downregulate N-cadherin expression.

Inhibitors of the lipogenic enzyme fatty acid synthase (FASN) have attracted much attention in the last decade as potential targeted cancer therapies. However, little is known about the molecular determinants of cancer cell sensitivity to FASN inhibitors (FASNis), which is a major roadblock to their therapeutic application. Here, we find that pharmacological starvation of endogenously produced FAs is a previously unrecognized metabolic stress that heightens mitochondrial apoptotic priming and favors cell death induction by BH3 mimetic inhibitors. Evaluation of the death decision circuits controlled by the BCL-2 family of proteins revealed that FASN inhibition is accompanied by the upregulation of the pro-death BH3-only proteins BIM, PUMA, and NOXA. Cell death triggered by FASN inhibition, which causally involves a palmitate/NADPH-related redox imbalance, is markedly diminished by concurrent loss of BIM or PUMA, suggesting that FASN activity controls cancer cell survival by fine-tuning the BH3 only proteins-dependent mitochondrial threshold for apoptosis. FASN inhibition results in a heightened mitochondrial apoptosis priming, shifting cells toward a primed-for-death state “addicted” to the anti-apoptotic protein BCL-2. Accordingly, co-administration of a FASNi synergistically augments the apoptosis-inducing activity of the dual BCL-XL/BCL-2 inhibitor ABT-263 (navitoclax) and the BCL-2 specific BH3-mimetic ABT-199 (venetoclax). FASN inhibition, however, fails to sensitize breast cancer cells to MCL-1- and BCL-XL-selective inhibitors such as S63845 and A1331852. A human breast cancer xenograft model evidenced that oral administration of the only clinically available FASNi drastically sensitizes FASN-addicted breast tumors to ineffective single-agents navitoclax and venetoclax in vivo. In summary, a novel FASN-driven facet of the mitochondrial priming mechanistically links the redox-buffering mechanism of FASN activity to the intrinsic apoptotic threshold in breast cancer cells. Combining next-generation FASNis with BCL-2-specific BH3 mimetics that directly activate the apoptotic machinery might generate more potent and longer-lasting antitumor responses in a clinical setting., The authors would like to thank Dr. Kenneth McCreath for editorial support. This work was supported by the NIH National Cancer Institute Grants R01 CA116623 (to Ruth Lupu) and R01 CA166741 (to Scott H. Kaufmann) and by the U.S. Department of Defense (DOD)-Breakthrough 3 Grants BC151072 and BC151072P1 (to Ruth Lupu). Work in the Menendez laboratory is supported by the Spanish Ministry of Science and Innovation (Grants SAF2016-80639-P and PID2019-10455GB-I00, Plan Nacional de l + D + I, founded by the European Regional Development Fund, Spain) and by an unrestricted research grant from the Fundació Oncolliga Girona (Lliga catalana d’ajuda al malalt de càncer, Girona). Joan Montero acknowledges support from the Ramon y Cajal Programme, Ministerio de Economía y Competitividad (RYC-2015-18357) and the Spanish National Plan “Retos Investigación” I + D + I (RTI2018-094533-A-I00) from the Ministerio de Ciencia, Innovación y Universidades. Elisabet Cuyàs holds a research contract “Miguel Servet” (CP20/00003) from the Instituto de Salud Carlos III, Spanish Ministry of Science and Innovation (Spain). All authors have read and agreed to the published version of the manuscript.

Simple Summary During the last decade, cold atmospheric plasmas (CAP) have been broadly investigated for their therapeutic effect against cancer. CAP sources can be used to treat liquid media, thereby generating plasma-conditioned liquids (PCL). PCL represent a very interesting alternative to direct CAP treatment, because they may allow treatment of malignant tumors located in inner organs of the body by means of an injection, thus avoiding multiple surgeries. Although research on this therapy is still in its early stage, PCL have already demonstrated their potential anticancer effect in different types of cancer in vivo. This review gathers the existing literature involving PCL treatments in vivo, highlighting the differences between the approaches undertaken and the need for establishing standardized protocols in order to better understand the effects of PCL-based therapies in vivo. Abstract Plasma-conditioned liquids (PCL) are gaining increasing attention in the medical field, especially in oncology, and translation to the clinics is advancing on a good path. This emerging technology involving cold plasmas has great potential as a therapeutic approach in cancer diseases, as PCL have been shown to selectively kill cancer cells by triggering apoptotic mechanisms without damaging healthy cells. In this context, PCL can be injected near the tumor or intratumorally, thereby allowing the treatment of malignant tumors located in internal organs that are not accessible for direct cold atmospheric plasma (CAP) treatment. Therefore, PCL constitutes a very interesting and minimally invasive alternative to direct CAP treatment in cancer therapy, avoiding surgeries and allowing multiple local administrations. As the field advances, it is progressively moving to the evaluation of the therapeutic effects of PCL in in vivo scenarios. Exciting developments are pushing forward the clinical translation of this novel therapy. However, there is still room for research, as the quantification and identification of reactive oxygen and nitrogen species (RONS) in in vivo conditions is not yet clarified, dosage regimens are highly variable among studies, and other more relevant in vivo models could be used. In this context, this work aims to present a critical review of the state of the field of PCL as anticancer agents applied in in vivo studies.

Microenvironment contributes to follicular lymphoma (FL) pathogenesis and impacts survival with macrophages playing a controversial role. In the present study, using FL primary samples and HK follicular dendritic cells (FDC) to mimic the germinal center, together with mouse models, we have analyzed the three-way crosstalk of FL-FDC-macrophages and derived therapeutic opportunities. Ex vivo primary FL-FDC co-cultures (n = 19) and in vivo mouse co-xenografts demonstrated that FL-FDC crosstalk favors tumor growth and, via the secretion of CCL2 and CSF-1, promotes monocyte recruitment, differentiation, and polarization towards an M2-like protumoral phenotype. Moreover, FL-M2 co-cultures displayed enhanced angiogenesis, dissemination, and immunosuppression. Analysis of the CSF-1/CSF-1R pathway uncovered that CSF-1 was significantly higher in serum from grade 3A FL patients, and that high CSF-1R expression in FL biopsies correlated with grade 3A, reduced overall survival and risk of transformation. Furthermore, CSF-1R inhibition with pexidartinib (PLX3397) preferentially affected M2-macrophage viability and polarization program disrupting FL-M2 positive crosstalk. In vivo CSF1-R inhibition caused M2 reduction and repolarization towards M1 macrophages and antitumor effect cooperating with anti-CD20 rituximab. In summary, these results support the role of macrophages in FL pathogenesis and indicate that CSF-1R may be a relevant prognostic factor and a novel therapeutic target cooperating with anti-CD20 immunotherapy. Grants that contributed to this work included: Gilead Sciences Research Funded Agreement, Spanish Ministry of Economy and Competitiveness & European Regional Development Fund (ERDF) “Una manera de hacer Europa” for SAF2014/57708R and SAF2017/ 88275R to PP-G and MCC, RTI2018-094584-B-I00 to DC, CIBERONC (CB16/12/00334 and CB16/12/00225), and finally Generalitat de Catalunya support for AGAUR 2017SGR1009 to DC

Thymidine kinase expressing human adipose mesenchymal stem cells (TK-hAMSCs) in combination with ganciclovir (GCV) are an effective platform for antitumor bystander therapy in mice models. However, this strategy requires multiple TK-hAMSCs administrations and a substantial number of cells. Therefore, for clinical translation, it is necessary to find a biocompatible scaffold providing TK-hAMSCs retention in the implantation site against their rapid wash-out. We have developed a microtissue (MT) composed by TKhAMSCs and a scaffold made of polylactic acid microparticles and cell-derived extracellular matrix deposited by hAMSCs. The efficacy of these MTs as vehicles for TK-hAMSCs/GCV bystander therapy was evaluated in a rodent model of human prostate cancer. Subcutaneously implanted MTs were integrated in the surrounding tissue, allowing neovascularization and maintenance of TK-hAMSCs viability. Furthermore, MTs implanted beside tumors allowed TK-hAMSCs migration towards tumor cells and, after GCV administration, inhibited tumor growth. These results indicate that TK-hAMSCs-MTs are promising cell reservoirs for clinical use of therapeutic MSCs in bystander therapies., This work was supported by the Severo Ochoa Program for Centers of Excellence in R&D 2016–2019, the European Commission-ERANET (nAngioderm JTC2018-103), the Spanish network of cell therapy (TERCEL), the Spanish Ministry of Science, Innovation and Universities (MAT2015-68906-R), the Spanish State Research Agency (AEI) and the European Regional Development Fund (FEDER) grants. BBF acknowledge finanical support through the BEST Postdoctoral Program, funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie (grant agreement no. 712754) and from the Spanish Ministry of Economy and Competitiveness under Severo Ochoa grants (SEV-2014-0425 and CEX2018-000789-S). GRS is thankful to the Spanish Ministry of Economy, Industry and Competitiveness for his fellowship (BES-2016-077182). The authors specially thank Dr. Josep Roca, from Delfos hospital (Dr. Roca I Noguera aesthetic surgery team), for the kind donation of liposuction for hAMSCs preparation; and to the Services of cell culture (IQAC-CISC), animal care (IQAC-CSIC) and cell sorting (CCiT-University of Barcelona) for their technician and specialized support.

Simple Summary The use of targeted drugs has brought about the development of new imaging techniques which are able to assess in vivo processes and changes in vascularization parameters can be captured as part of the antitumor response to antiangiogenic therapies. This pilot study (IMPACT trial, NCT02316327) aimed to explore the capacity of Perfusion-Computed Tomography (pCT) to detect early changes in tumor vascularization in non-small cell lung cancer (NSCLC) patients treated with an antiangiogenic-based therapy. Our results confirm the feasibility of pCT to capture early changes in tumor vasculature and suggest the potential of blood volume (BV) to early identify differential tumor responses to antiangiogenic therapy. Abstract Background: The role of perfusion computed tomography (pCT) in detecting changes in tumor vascularization as part of a response to antiangiogenic therapy in non-small cell lung cancer (NSCLC) remains unclear. Methods: In this prospective pilot study (IMPACT trial, NCT02316327), we aimed to determine the ability of pCT to detect early changes in blood flow (BF), blood volume (BV), and permeability (PMB), and to explore whether these changes could predict the response at day +42 in patients with advanced, treatment-naive, non-squamous NSCLC treated with cisplatin and gemcitabine plus bevacizumab. Results: All of the perfusion parameters showed a consistent decrease during the course of treatment. The BV difference between baseline and early assessment was significant (p = 0.013), whereas all perfusion parameters showed significant differences between baseline and day +42 (p = 0.003, p = 0.049, and p = 0.002, respectively). Among the 16 patients evaluable for efficacy, a significant decline in BV at day +7 from baseline was observed in tumors with no response (p = 0.0418). Conclusions: Our results confirm that pCT can capture early changes in tumor vasculature. A substantial early decline of BV from baseline might identify tumors less likely responsive to antiangiogenic-drugs.

Molecular profiling of personal cancer genomes, and the identification of actionable vulnerabilities and drug-response biomarkers, are the basis of precision oncology. Tumors often present several driver alterations that might be connected by cross-talk and feedback mechanisms, making it difficult to mark single oncogenic variations as reliable predictors of therapeutic outcome. In the current work, we uncover and exploit driver alteration co-occurrence patterns from a recently published in vivo screening in patient-derived xenografts (PDXs), including 187 tumors and 53 drugs. For each treatment, we compare the mutational profiles of sensitive and resistant PDXs to statistically define Driver Co-Occurrence (DCO) networks, which capture both genomic structure and putative oncogenic synergy. We then use the DCO networks to train classifiers that can prioritize, among the available options, the best possible treatment for each tumor based on its oncogenomic profile. In a cross-validation setting, our drug-response models are able to correctly predict 66% of sensitive and 77% of resistant drug-tumor pairs, based on tumor growth variation. Perhaps more interesting, our models are applicable to several tumor types and drug classes for which no biomarker has yet been described. Additionally, we experimentally validated the performance of our models on 15 new tumor samples engrafted in mice, achieving an overall accuracy of 75%. Finally, we adapted our strategy to derive drug-response models from continuous clinical outcome measures, such as progression free survival, which better represent the data acquired during routine clinical practice and in clinical trials. We believe that the computational framework presented here could be incorporated into the design of adaptive clinical trials, revealing unexpected connections between oncogenic alterations and increasing the clinical impact of genomic profiling.

Epidermal growth factor receptor (EGFR) tyrosine kinases inhibitors (TKIs) are effective therapies for non-small cell lung cancer (NSCLC) patients whose tumors harbor an EGFR activating mutation. However, this treatment is not curative due to primary and secondary resistance such as T790M mutation in exon 20. Recently, activation of transducer and activator of transcription 3 (STAT3) in NSCLC appeared as an alternative resistance mechanism allowing cancer cells to elude the EGFR signaling. Overexpression of fatty acid synthase (FASN), a multifunctional enzyme essential for endogenous lipogenesis, has been related to resistance and the regulation of the EGFR/Jak2/STAT signaling pathways. Using EGFR mutated (EGFRm) NSCLC sensitive and EGFR TKIs&rsquo, resistant models (Gefitinib Resistant, GR) we studied the role of the natural polyphenolic anti-FASN compound (&minus, )-epigallocatechin-3-gallate (EGCG), and its derivative G28 to overcome EGFR TKIs&rsquo, resistance. We show that G28&rsquo, s cytotoxicity is independent of TKIs&rsquo, resistance mechanisms displaying synergistic effects in combination with gefitinib and osimertinib in the resistant T790M negative (T790M&minus, ) model and showing a reduction of activated EGFR and STAT3 in T790M positive (T790M+) models. Our results provide the bases for further investigation of G28 in combination with TKIs to overcome the EGFR TKI resistance in NSCLC.

Publisher's version (útgefin grein), Sex differences have been observed in multiple facets of cancer epidemiology, treatment and biology, and in most cancers outside the sex organs. Efforts to link these clinical differences to specific molecular features have focused on somatic mutations within the coding regions of the genome. Here we report a pan-cancer analysis of sex differences in whole genomes of 1983 tumours of 28 subtypes as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium. We both confirm the results of exome studies, and also uncover previously undescribed sex differences. These include sex-biases in coding and non-coding cancer drivers, mutation prevalence and strikingly, in mutational signatures related to underlying mutational processes. These results underline the pervasiveness of molecular sex differences and strengthen the call for increased consideration of sex in molecular cancer research., We thank all the members of the Boutros lab for insightful discussions. This study was conducted with the support of the Ontario Institute for Cancer Research to P.C.B. through funding provided by the Government of Ontario. This work was supported by the Discovery Frontiers: Advancing Big Data Science in Genomics Research program, which is jointly funded by the Natural Sciences and Engineering Research Council (NSERC) of Canada, the Canadian Institutes of Health Research (CIHR), Genome Canada and the Canada Foundation for Innovation (CFI). P.C.B. was supported by a Terry Fox Research Institute New Investigator Award and a CIHR New Investigator Award. This work was supported by an NSERC Discovery grant and by Canadian Institutes of Health Research, grant #SVB-145586, to P.C.B. This work was supported by the NIH/NCI under award number P30CA016042 and an operating grant from the National Cancer Institute Early Detection Research Network (1U01CA214194-01). We acknowledge the contributions of the many clinical networks across ICGC and TCGA who provided samples and data to the PCAWG Consortium, and the contributions of the Technical Working Group and the Germline Working Group of the PCAWG Consortium for collation, realignment and harmonised variant calling of the cancer genomes used in this study. We thank the patients and their families for their participation in the individual ICGC and TCGA projects.

Metabolic reprogramming, carried out by cancer cells to rapidly adapt to stress such as hypoxia and limited nutrient conditions, is an emerging concepts in tumor biology, and is now recognized as one of the hallmarks of cancer. In contrast with conventional views, based on the classical Warburg effect, these metabolic alterations require fully functional mitochondria and finely-tuned regulations of their activity. In turn, the reciprocal regulation of the metabolic adaptations of cancer cells and the microenvironment critically influence disease progression and response to therapy. This is also realized through the function of specific stress-adaptive proteins, which are able to relieve oxidative stress, inhibit apoptosis, and facilitate the switch between metabolic pathways. Among these, the molecular chaperone tumor necrosis factor receptor associated protein 1 (TRAP1), the most abundant heat shock protein 90 (HSP90) family member in mitochondria, is particularly relevant because of its role as an oncogene or a tumor suppressor, depending on the metabolic features of the specific tumor. This review highlights the interplay between metabolic reprogramming and cancer progression, and the role of mitochondrial activity and oxidative stress in this setting, examining the possibility of targeting pathways of energy metabolism as a therapeutic strategy to overcome drug resistance, with particular emphasis on natural compounds and inhibitors of mitochondrial HSP90s.

// Elisabet Cuyas 1,2,* , Sara Verdura 1,2,* , Salvador Fernandez-Arroyo 3,4 , Joaquim Bosch-Barrera 5 , Begona Martin-Castillo 6 , Jorge Joven 3,4 and Javier A. Menendez 1,2 1 Metabolism and Cancer Group, Program Against Cancer Therapeutic Resistance, Catalan Institute of Oncology, Girona, Spain 2 Molecular Oncology Group, Girona Biomedical Research Institute (IDIBGI), Girona, Spain 3 Unitat de Recerca Biomedica, Hospital Universitari de Sant Joan, Institut d’Investigacio Sanitaria Pere Virgili, Universitat Rovira i Virgili, Reus, Catalonia, Spain 4 Campus of International Excellence Southern Catalonia, Tarragona, Catalonia, Spain 5 Medical Oncology, Catalan Institute of Oncology, Girona, Catalonia, Spain 6 Unit of Clinical Research, Catalan Institute of Oncology, Girona, Spain * These authors have contributed equally to this work Correspondence to: Javier A. Menendez, email: // Keywords : cancer stem cells; metabolic maps; tumor heterogeneity Received : July 21, 2017 Accepted : September 29, 2017 Published : October 15, 2017 Abstract Personalized cancer medicine based on the analysis of tumors en masse is limited by tumor heterogeneity, which has become a major obstacle to effective cancer treatment. Cancer stem cells (CSC) are emerging as key drivers of inter- and intratumoral heterogeneity. CSC have unique metabolic dependencies that are required not only for specific bioenergetic/biosynthetic demands but also for sustaining their operational epigenetic traits, i.e. self-renewal, tumor-initiation, and plasticity. Given that the metabolome is the final downstream product of all the –omic layers and, therefore, most representative of the biological phenotype, we here propose that a novel approach to better understand the complexity of tumor heterogeneity is by mapping and cataloging small numbers of CSC metabolomic phenotypes. The narrower metabolomic diversity of CSC states could be employed to reduce multidimensional tumor heterogeneity into dynamic models of fewer actionable sub-phenotypes. The identification of the driver nodes that are used differentially by CSC states to metabolically regulate self-renewal and tumor initation and escape chemotherapy might open new preventive and therapeutic avenues. The mapping of CSC metabolomic states could become a pioneering strategy to reduce the dimensionality of tumor heterogeneity and improve our ability to examine changes in tumor cell populations for cancer detection, prognosis, prediction/monitoring of therapy response, and detection of therapy resistance and recurrent disease. The identification of driver metabolites and metabolic nodes accounting for a large amount of variance within the CSC metabolomic sub-phenotypes might offer new unforeseen opportunities for reducing and exploiting tumor heterogeneity via metabolic targeting of CSC.

The development of a single immuno-metabolic adjuvant capable of modulating, in the appropriate direction and intensity, the complex antagonistic and symbiotic interplays between tumor cells, immune cells, and the gut microbiota may appear pharmacologically implausible. Metformin might help solve this conundrum and beneficially impact the state of cancer-immune system interactions Work in the Menendez laboratory is supported by the Spanish Ministry of Science and Innovation (Grant SAF2016-80639-P, Plan Nacional de l +D+I, founded by the European Regional Development Fund, Spain) and by an unrestricted research grant from the Fundació Oncolliga Girona (Lliga catalana d’ajuda al malalt de càncer, Girona)

Homocisteïna; Càncer de mama; Dieta cetogènica Homocisteína; Cáncer de mama; Dieta cetogénica Homocysteine; Breast cancer; Ketogenic diet Certain dietary interventions might improve the therapeutic index of cancer treatments. An alternative to the “drug plus diet” approach is the pharmacological reproduction of the metabolic traits of such diets. Here we explored the impact of adding metformin to an established therapeutic regimen on the systemic host metabolism of cancer patients. A panel of 11 serum metabolites including markers of mitochondrial function and intermediates/products of folate-dependent one-carbon metabolism were measured in paired baseline and post-treatment sera obtained from HER2-positive breast cancer patients randomized to receive either metformin combined with neoadjuvant chemotherapy and trastuzumab or an equivalent regimen without metformin. Metabolite profiles revealed a significant increase of the ketone body β-hydroxybutyrate and of the TCA intermediate α-ketoglutarate in the metformin-containing arm. A significant relationship was found between the follow-up levels of homocysteine and the ability of treatment arms to achieve a pathological complete response (pCR). In the metformin-containing arm, patients with significant elevations of homocysteine tended to have a higher probability of pCR. The addition of metformin to an established anti-cancer therapeutic regimen causes a fasting-mimicking modification of systemic host metabolism. Circulating homocysteine could be explored as a clinical pharmacodynamic biomarker linking the antifolate-like activity of metformin and biological tumor response. This work was supported by grants from the Ministerio de Sanidad, Servicios Sociales e Igualdad (EC10-125, Ayudas para el Fomento de la Investigación Clínica Independiente to Begoña Martin-Castillo). Work in the Menendez laboratory is supported by the Ministerio de Ciencia e Innovación (Grant SAF2016-80639-P, Plan Nacional de l+D+I, founded by the European Regional Development Fund [EU FEDER], Spain) and by an unrestricted research grant from the Fundació Oncolliga Girona (Lliga catalana d’ajuda al malalt de càncer, Girona).

Acquired resistance to trastuzumab is a major clinical problem in the treatment of HER2-positive (HER2+) breast cancer patients. The selection of trastuzumab-resistant patients is a great challenge of precision oncology. The aim of this study was to identify novel epigenetic biomarkers associated to trastuzumab resistance in HER2+ BC patients. Methods: We performed a genome-wide DNA methylation (450K array) and a transcriptomic analysis (RNA-Seq) comparing trastuzumab-sensitive (SK) and trastuzumab-resistant (SKTR) HER2+ human breast cancer cell models. The methylation and expression levels of candidate genes were validated by bisulfite pyrosequencing and qRT-PCR, respectively. Functional assays were conducted in the SK and SKTR models by gene silencing and overexpression. Methylation analysis in 24 HER2+ human BC samples with complete response or non-response to trastuzumab-based treatment was conducted by bisulfite pyrosequencing. Results: Epigenomic and transcriptomic analysis revealed the consistent hypermethylation and downregulation of TGFBI, CXCL2, and SLC38A1 genes in association with trastuzumab resistance. The DNA methylation and expression levels of these genes were validated in both sensitive and resistant models analyzed. Of the genes, TGFBI presented the highest hypermethylation-associated silencing both at the transcriptional and protein level. Ectopic expression of TGFBI in the SKTR model suggest an increased sensitivity to trastuzumab treatment. In primary tumors, TGFBI hypermethylation was significantly associated with trastuzumab resistance in HER2+ breast cancer patients. Conclusions: Our results suggest for the first time an association between the epigenetic silencing of TGFBI by DNA methylation and trastuzumab resistance in HER2+ cell models. These results provide the basis for further clinical studies to validate the hypermethylation of TGFBI promoter as a biomarker of trastuzumab resistance in HER2+ breast cancer patients This work was supported in part by the Spanish Instituto de Salud Carlos III (ISCIII; FIS PI11/00692 and PI14/00329; to’ T. Puig), Fundacion Ramon Areces (to T. Puig), the support of Catalonian Government (2017SGR00385), and La Marató de TV3 (20131530, TPuig), financial support was from the University of Girona (MPCUdG2016/036), and the University of Girona and La Caixa Foundation awarded S. Palomeras with a predoctoral grant. A.D.-L. is funded by a “Juan Rodés” contract (JR17/00016) from ISCIII. Ana B Crujeiras is funded by a research contract “Miguel Servet” (CP17/00088) from the ISCIII

En los últimos años, la medicina de plasma ha sido un tema emergente de estudio e investigación en distintas áreas (esterilización, curación de heridas, tratamientos dentales), y muy especialmente en el área de terapias de cáncer. Estudios recientes han demostrado que las especies reactivas de oxígeno y nitrógeno que genera el plasma tienen un efecto selectivo y citotóxico en las células cancerígenas, sin afectar a las células sanas. Se han demostrado efectos letales del plasma en más de 20 líneas celulares in vitro e in vivo, y recientemente, en el grupo de investigación BBT, en cáncer de hueso. El tratamiento plasma puede efectuarse de forma directa, aplicando directamente el plasma sobre el tumor o las células, de modo que todos los componentes del plasma interaccionan con el material biológico; o de forma indirecta, poniendo en contacto con el tumor o las células un medio líquido previamente tratado con plasma, de manera que únicamente las especies reactivas que se generan en el medio líquido interaccionan con el material biológico. El objetivo principal de este proyecto es investigar los efectos directos e indirectos del plasma a presión atmosférica sobre dos líneas celulares de osteosarcoma, y estudiar dichos efectos tanto en 2D (células sembradas en placas de cultivo) como en 3D, haciendo crecer las células tumorales en andamios tridimensionales para mimetizar mejor el microambiente tumoral. Para ello, se han caracterizado las especies reactivas (H2O2, NO2-, y especies de tiempo de vida corto) generadas por el plasma atmosférico en medio de cultivo a diferentes tiempos de tratamiento. La concentración de especies en el medio aumenta progresivamente con el tiempo de tratamiento. Para realizar el estudio del efecto del plasma en 3D, se han sintetizado nanopartículas de hidroxiapatita para incorporarlas a andamios de colágeno. Se ha analizado la degradación y capacidad de absorción de líquidos de los andamios con el fin de comprobar su aptitud para la siembra de células. Posteriormente se ha estudiado los efectos directos e indirectos del plasma sobre las células sembradas en los andamios. Se ha comprobado que la concentración de especies reactivas que se generan es proporcional al tiempo de tratamiento, y en contraste con trabajos anteriores, se ha observado que la viabilidad de las células tratadas por plasma de manera indirecta no se ve afectada. Esto es debido a que se ha empleado medio de cultivo con piruvato, que actúa como scavenger de los peróxidos. Con el tratamiento directo sí que hay un ligero efecto citotóxico en las células, tanto en 2D como en 3D. Objectius de Desenvolupament Sostenible::3 - Salut i Benestar

Cancer is a worldwide known disease that affects a big part of the population, reason why it’s being studied nowadays in biomedical research. The tyrosine kinase receptors family ErbB is highly studied in cancer therapy, due to its implication in cell proliferation and differentiation pathways. The epidermal growth factor (EGF) is a ligand of this family that binds to its receptor EGFR causing a dimerization and consequently autophosphorylation, driving to the activation of the signaling path. Targeted cancer therapy it’s about guiding a drug to the tumoral cells through determined proteins (usually receptors) involved in the tumoral process. The idea of this project is to address the attack to the epidermal growth factor receptor (EGFR), which is involved in the uncontrolled massive cell proliferation characteristic of this disease, and lock it with a derivative of its own EGF ligand. To convert this ligand into a blocker agent is necessary to do some modifications to the residues that composes it, with the intention of maintain the natural affinity of this peptide to its receptor but avoiding that this causes dimerization, eluding the activation of the cell growth pathway. This final degree work is focused in the study of the natural interaction between the EGF ligand and its EGFR receptor, comparing it with the truncated model (EGFt) done by Panosa et al. in 2013. By molecular dynamics the trajectories of both models are generated and after analyzed by Amber16. Finally, docking of proteins is done by ClusPro 2.0 to foresee the interaction that would be between the modified ligands (EGFm) and the receptor

Gramicidin (GA), a very hydrophobic pentadecapeptide with important biological activities (i.e. in addition to its well-known antimicrobial and antibiotic activities, GA has been recently identified as a potent therapeutic agent against different carcinomas), has been loaded by electrospraying in poly(tetramethylene succinate) (PE44), a biodegradable and biocompatible aliphatic polyester. Microspheres (average diameter: 5.0 ± 0.7 μm) were successfully obtained from the mixture of GA and PE44 solutions in ethanol and chloroform, respectively. The loading of the peptide, which has been proved by FTIR and X-ray photoelectron spectroscopies, essentially occurred at the surface of the microspheres, as was reflected by scanning electron microscopy micrographs and atomic force microscopy phase images. In spite of this, the thermal stability of the polyester matrix remained essentially unaltered, even though the wettability decreased. The release of GA in phosphate buffer saline (PBS) was limited by the very low solubility of the peptide in aqueous solution, a fast burst effect followed by the establishment of equilibrium after 5 days of being observed in this hydrophilic environment. The release behaviour was very different when the hydrophilicity of the medium was reduced by adding ethanol. In this case, a very fast but sustained release was identified during the first few hours. On the other hand, biological tests have demonstrated that GA retains its antimicrobial activity after loading and does not alter the biocompatibility of PE44. Our results prove that, despite its hydrophobicity and relatively large number of residues, the loading of GA in a polymeric matrix represents an alternative strategy for the release of this versatile peptide in cancer therapy., Authors thank supports from MINECO and FEDER (MAT2015-69367-R, MAT2015-69547-R and CTQ2013-40855-R) and Gobierno de Aragón – Fondo Social Europeo (research group E40). Support for the research of C.A. was received through the prize “ICREA Academia” for excellence in research funded by the Generalitat de Catalunya.

Atmospheric pressure plasma jets (APPJ) have great potential in wound healing, bacterial disinfection and in cancer therapy. Recent studies pointed out that hydrogels can be used as screens during APPJ treatment, or even be used as reservoirs for reactive oxygen and nitrogen species generated by APPJ in liquids. Thus, novel applications are emerging for hydrogels which deserve fundamental exploration of the possible modifications undergone by the polymers in solution due to the reactivity with plasmas. Here we investigate the possible modifications occurred by APPJ treatment of an amphiphilic poly(ethylene oxide)-based triblock copolymer (tPEO) photo-crosslinkable hydrogel. While APPJ treatments lead to a certain degradation of the self-assembly of the polymeric chains at low concentrations (2 g/L), the polymeric chains are unaffected by APPJ and the hydrogel forming ability is kept. APPJ treatments induced a pre-crosslinking of the network with an increase of the mechanical properties of the hydrogel. Overall, the small modifications induced allow thinking of polymer solutions with hydrogel forming ability a new platform for several applications related to plasma medicine, and thus, with potential in different therapies.

This work was supported by the Agencia Estatal de Investigación (AEI)[MICINN/Fondo Europeo de Desarrollo Regional (FEDER) (RTC2016-4603-1 to EntreChem SL and SAF-2016-75286-R toR.R.), ISC III/FEDER [Miguel Servet Program] (CPII16/00049 to R.R.), Sara Borrell Program (CD16/00103 toS-T.M.) and Consorcio CIBERONC (CB16/12/00390)] and the Plan de Ciencia Tecnología e Innovación del Principado de Asturias (GRUPIN14-003). We want to particularly acknowledge for its collaboration, the Principado de Asturias BioBank (PT17/0015/0023),financed jointly by Servicio de Salud del Principado de Asturias, Instituto de Salud Carlos III and Fundación Bancaria Cajastur and integrated in the Spanish National Biobanks Network., Estupiñan, O., Santos, L., Rodriguez, A., Fernandez-Nevado, L., Costales, P., Perez-Escuredo, J., Hermosilla, M.A., Oro, P., Rey, V., Tornin, J., Allonca, E., Fernandez-Garcia, M.T., Alvarez-Fernandez, C., Braña, A., Astudillo, A., Menendez, S.T., Moris, F., Rodriguez, R.

KRAS mutations are an established predictor of lack of response to EGFR-targeted therapies in patients with metastatic colorectal cancer (mCRC). However, little is known about the role of the rarer NRAS mutations as a mechanism of primary resistance to the anti-EGFR monoclonal antibody cetuximab in wild-type KRAS mCRC. Using isogenic mCRC cells with a heterozygous knock-in of the NRAS activating mutation Q61K, we aimed to elucidate the mechanism(s) by which mutant NRAS blocks cetuximab from inhibiting mCRC growth. NRASQ61K/+ cells were refractory to cetuximab-induced growth inhibition. Pathway-oriented proteome profiling revealed that cetuximab-unresponsive ERK1/2 phosphorylation was the sole biomarker distinguishing cetuximab-refractory NRASQ61K/+ from cetuximab-sensitive NRAS+/+ cells. We therefore employed four representative MEK1/2 inhibitors (binimetinib, trametinib, selumetinib, and pimasertib) to evaluate the therapeutic value of MEK/ERK signaling in cetuximab-refractory NRAS mutation-induced mCRC. Co-treatment with an ineffective dose of cetuximab augmented, up to more than 1,300-fold, the cytotoxic effects of pimasertib against NRASQ61K/+ cells. Simultaneous combination of MEK1/2 inhibitors with cetuximab resulted in extremely high and dose-dependent synthetic lethal effects, which were executed, at least in part, by exacerbated apoptotic cell death. Dynamic monitoring of real-time cell growth rates confirmed that cetuximab synergistically sensitized NRASQ61K/+ cellsto MEK1/2 inhibition. Our discovery of a synthetic lethal interaction of cetuximab in combination with MEK1/2 inhibition for the NRAS mutant subgroup of mCRC underscores the importance of therapeutic intervention both in the MEK-ERK and EGFR pathways to achieve maximal therapeutic efficacy against NRAS-mutant mCRC tumors.

Background Ribonucleases are promising agents for use in anticancer therapy. Among the different ribonucleases described to be cytotoxic, a paradigmatic example is onconase which manifests cytotoxic and cytostatic effects, presents synergism with several kinds of anticancer drugs and is currently in phase II/III of its clinical trial as an anticancer drug against different types of cancer. The mechanism of cytotoxicity of PE5, a variant of human pancreatic ribonuclease carrying a nuclear localization signal, has been investigated and compared to that of onconase. Methods Cytotoxicity was measured by the MTT method and by the tripan blue exclusion assay. Apoptosis was assessed by flow cytometry, caspase enzymatic detection and confocal microscopy. Cell cycle phase analysis was performed by flow cytometry. The expression of different proteins was analyzed by western blot. Results We show that the cytotoxicity of PE5 is produced through apoptosis, that it does not require the proapoptotic activity of p53 and is not prevented by the multiple drug resistance phenotype. We also show that PE5 and onconase induce cell death at the same extent although the latter is also able to arrest the cell growth. We have compared the cytotoxic effects of both ribonucleases in the NCI/ADR-RES cell line by measuring their effects on the cell cycle, on the activation of different caspases and on the expression of different apoptosis- and cell cycle-related proteins. PE5 increases the number of cells in S and G2/M cell cycle phases, which is accompanied by the increased expression of cyclin E and p21WAF1/CIP1 together with the underphosphorylation of p46 forms of JNK. Citotoxicity of onconase in this cell line does not alter the cell cycle phase distribution and it is accompanied by a decreased expression of XIAP Conclusions We conclude that PE5 kills the cells through apoptosis associated with the p21WAF1/CIP1 induction and the inactivation of JNK. This mechanism is significantly different from that found for onconase.

Background Methotrexate is a chemotherapeutic drug that is used in therapy of a wide variety of cancers. The efficiency of treatment with this drug is compromised by the appearance of resistance. Combination treatments of MTX with other drugs that could modulate the expression of genes involved in MTX resistance would be an adequate strategy to prevent the development of this resistance. Methods The differential expression pattern between sensitive and MTX-resistant cells was determined by whole human genome microarrays and analyzed with the GeneSpring GX software package. A global comparison of all the studied cell lines was performed in order to find out differentially expressed genes in the majority of the MTX-resistant cells. S100A4 mRNA and protein levels were determined by RT-Real-Time PCR and Western blot, respectively. Functional validations of S100A4 were performed either by transfection of an expression vector for S100A4 or a siRNA against S100A4. Transfection of an expression vector encoding for β-catenin was used to inquire for the possible transcriptional regulation of S100A4 through the Wnt pathway. Results S100A4 is overexpressed in five out of the seven MTX-resistant cell lines studied. Ectopic overexpression of this gene in HT29 sensitive cells augmented both the intracellular and extracellular S100A4 protein levels and caused desensitization toward MTX. siRNA against S100A4 decreased the levels of this protein and caused a chemosensitization in combined treatments with MTX. β-catenin overexpression experiments support a possible involvement of the Wnt signaling pathway in S100A4 transcriptional regulation in HT29 cells. Conclusions S100A4 is overexpressed in many MTX-resistant cells. S100A4 overexpression decreases the sensitivity of HT29 colon cancer human cells to MTX, whereas its knockdown causes chemosensitization toward MTX. Both approaches highlight a role for S100A4 in MTX resistance.

The Epidermal Growth Factor Receptor (EGFR) is a glycoprotein that belongs to the family of tyrosine kinase receptors and that it is usually altered in tumor cells. It has been reported that its inhibition decreases tumor proliferation and thus, it is an attractive target for antitumor therapies. There are currently two main strategies of EGFR inhibition: monoclonal antibodies and tyrosine kinase inhibitors (TKI). Unfortunately, the appearance of resistance and other factors limit often the effectiveness of these treatments. In order to study how certain changes in EGFR can affect target therapies against this receptor, we must understand its conformational dynamics. EGFR activation requires the binding of two monomers that, induced by the presence of a ligand, allow then the interaction of the kinase domains and initiate the downstream signaling. However, the deregulation of this receptor may be modulated by several factors, such as the glycosylation of its ectodomine. Tumor progression has been related with an alteration of EGFR glycosylation and it has been observed that aberrant glycosylations may affect both the binding of the ligand and its activation. Accordingly, to study the overall effect of glycosylation on the response to EGFR directed treatments requires a more profound investigation. This is the objective of this Degree Final Project. Based on the existing bibliographic information, it is widely accepted that the site and type of glycosylation influence EGFR conformation and activity. In spite of the controversy in some studies, it seems that terminal fucosylation and sialylation decrease the activity of this receptor. In contrast, other glycosylations such as core fucosylation and Lewis epitopes, increase the activity of EGFR. Moreover, it seems that the alteration of glycosylation could have an effect on EGFR response to treatments. According to current information, EGFR glycosylation does not seem to alter the binding of antibodies to the extracellular domain. However, studies on another member of the EGFR family, HER2, show the opposite. The most relevant result obtained in this project is that extracellular glycosylation of EGFR indirectly affects intracellular sensitivity to TKI. Finally, the need to find alternative treatments to fight emerging resistance and the demand for higher treatment specificity opens an avenue for glycosylation itself as a potential novel target against cancer

Supplemental Digital Content is Available in the Text., Basal cell carcinoma (BCC) seems to originate from ultraviolet light-induced mutations involving the bulge or the outer sheath of the hair follicle cells. However, the etiopathogenic mechanisms involved in the development of these tumors in nonphotoexposed and in hairless areas remain unclear. The cytokeratin (CK) profile (including CK5/6, CK7, CK14, CK15, CK17, and CK19) from a series of different BCC subtypes developing in sun-exposed and non–sun-exposed areas, including hairless regions, was evaluated. The authors have observed that CK7 expression in BCC is associated with the anatomical localization of the tumor and its sun-exposition, but not with other factors such as histological subtype. The expression of this CK is higher in BCCs located in non–sun-exposed and nonhairy areas, such as the vulvar semimucosa and the nipple. Because CK7 is a marker of simple glandular epithelia, the authors suggest a glandular origin for BCCs located in hairless and nonphotoexposed areas.

The undecapeptide KKLFKKILKKL-NH2 (BP16) is a non-toxic cell-penetrating peptide (CPP) that is mainly internalized into cancer cells through a clathrin dependent endocytic mechanism and localizes in late endosomes. Moreover, this CPP is able to enhance the cellular uptake of chlorambucil (CLB) improving its cytotoxicity. In this work, we further explored the cell-penetrating properties of BP16 and those of its arginine analogue BP308. We investigated the influence on the cytotoxicity and on the cellular uptake of conjugating CLB at the N- or the C-terminal end of these undecapeptides. The effect of incorporating the cathepsin B-cleavable sequence Gly-Phe-Leu-Gly in CLB-BP16 and CLB-BP308 conjugates was also evaluated. The activity of CLB was significantly improved when conjugated at the N- or the C-terminus of BP16, or at the N-terminus of BP308. While CLB alone was not active (IC50 of 73.7 to >100 μM), the resulting conjugates displayed cytotoxic activity against CAPAN-1, MCF-7, PC-3, 1BR3G and SKMEL-28 cell lines with IC50 values ranging from 8.7 to 25.5 μM. These results were consistent with the internalization properties observed for the corresponding 5(6)-carboxyfluorescein-labeled conjugates. The presence of the tetrapeptide Gly-Phe-Leu-Gly at either the N- or the C-terminus of CLB-BP16 conjugates further increased the efficacy of CLB (IC50 of 3.6 to 16.2 μM), which could be attributed to its selective release in the lysosomal compartment. Enzymatic assays with cathepsin B showed the release of CLB-Gly-OH from these sequences within a short time. Therefore, the combination of BP16 with an enzymatic cleavable sequence can be used as a drug delivery system for the effective uptake and release of drugs in cancer cells This work was supported by Consolider Ingenio CSD/CSD2010-00065 from MICINN of Spain. We also thank the Catalan DIUE of the Generalitat de Catalunya (2009SGR637). X. R. acknowledges financial support from INNPLANTA project INP-2011-0059-PCT-420000-ACT1

Contains fulltext : 170338.pdf (Publisher’s version ) (Open Access) Background Patients with advanced urothelial carcinoma that progresses after platinum-based chemotherapy have a poor prognosis and limited treatment options. Methods In this open-label, international, phase 3 trial, we randomly assigned 542 patients with advanced urothelial cancer that recurred or progressed after platinum-based chemotherapy to receive pembrolizumab (a highly selective, humanized monoclonal IgG4kappa isotype antibody against programmed death 1 [PD-1]) at a dose of 200 mg every 3 weeks or the investigator's choice of chemotherapy with paclitaxel, docetaxel, or vinflunine. The coprimary end points were overall survival and progression-free survival, which were assessed among all patients and among patients who had a tumor PD-1 ligand (PD-L1) combined positive score (the percentage of PD-L1-expressing tumor and infiltrating immune cells relative to the total number of tumor cells) of 10% or more. Results The median overall survival in the total population was 10.3 months (95% confidence interval [CI], 8.0 to 11.8) in the pembrolizumab group, as compared with 7.4 months (95% CI, 6.1 to 8.3) in the chemotherapy group (hazard ratio for death, 0.73; 95% CI, 0.59 to 0.91; P=0.002). The median overall survival among patients who had a tumor PD-L1 combined positive score of 10% or more was 8.0 months (95% CI, 5.0 to 12.3) in the pembrolizumab group, as compared with 5.2 months (95% CI, 4.0 to 7.4) in the chemotherapy group (hazard ratio, 0.57; 95% CI, 0.37 to 0.88; P=0.005). There was no significant between-group difference in the duration of progression-free survival in the total population (hazard ratio for death or disease progression, 0.98; 95% CI, 0.81 to 1.19; P=0.42) or among patients who had a tumor PD-L1 combined positive score of 10% or more (hazard ratio, 0.89; 95% CI, 0.61 to 1.28; P=0.24). Fewer treatment-related adverse events of any grade were reported in the pembrolizumab group than in the chemotherapy group (60.9% vs. 90.2%); there were also fewer events of grade 3, 4, or 5 severity reported in the pembrolizumab group than in the chemotherapy group (15.0% vs. 49.4%). Conclusions Pembrolizumab was associated with significantly longer overall survival (by approximately 3 months) and with a lower rate of treatment-related adverse events than chemotherapy as second-line therapy for platinum-refractory advanced urothelial carcinoma. (Funded by Merck; KEYNOTE-045 ClinicalTrials.gov number, NCT02256436 .).

Systemic treatment of cancer requires tumour-selective therapies that eliminate cancer cells yet preserve healthy tissues from undesired damage. Tumoral transformation is associated with profound effects in translational reprogramming of gene expression, such that tumour-specific translational regulation presents an attractive possibility for generating oncoselective therapies. We recently discovered that mRNA translational control by cytoplasmic polyadenylation element-binding proteins (CPEBs) is reactivated in cancer. Here we present a novel approach to restrict genetic-engineered therapies to malignant tissues based on CPEB translational regulation of target mRNAs. We demonstrate that tumour reprogramming of CPEB-mediated mRNA stability and translational regulation modulates tumour-specific expression of viral proteins. For oncolytic adenoviruses, insertion of CPE regulatory sequences in the 3′-untranslated region of the E1A gene provides oncoselectivity, with full potency in cancer cells but attenuated in normal tissues. Our results demonstrate the potential of this strategy to improve oncolytic virus design and provide a framework for exploiting CPE-regulated transgenes for therapy., mRNA translational control by CPEBs is reactivated in cancer. Here, the authors show that the insertion of CPE regulatory sequences in the 3′ UTR of the E1A viral gene induces oncoselectivity, with oncolytic potency in cancer cells but attenuated toxicity in normal tissues.

Targeted therapy drugs, mainly those within the signal transduction inhibitors, are used more chronically than cytotoxic drugs and are metabolised by cytochrome P450 isozymes so patients are at high risk of having drug-drug interactions (DDI). Not only this, as the majority of them are given orally, new drug-drug interactions concerning gastrointestinal absorption can occur (e.g., with proton pump inhibitors). DDI can lead to changed systemic exposure, resulting in variations in drug response of the co-administered. In addition, concomitant ingestion of dietary supplements could also alter systemic exposure of drugs, thus leading to adverse drug reactions or loss of efficacy. In this review, we give an overview of the current existing data of known or suspected DDI between targeted therapy and other medicines. A review of package inserts was performed to identify drug-drug interactions for all targeted antineoplastic agents. Tertiary databases such as Lexicomp®, Drugs, Martindale, Facts and Comparisons®, and AHFS Drug Information were also referenced. This study covered 40 targeted antineoplastic agents (28 signal transduction inhibitors, 9 monoclonal antibodies and 3 other drugs, 2 monoclonal antibody conjugates and 1 fusion protein). Most of targeted therapy drugs are major CYP3A4 substrates with P-gp playing an important role in disposition too. Thus, there is a very common thread here that these agents will likely be sensitive victims to strong CYP3A4/P-gp inhibitors and inducers. It is essential that health care providers monitor patients for potential DDI to avoid a loss in efficacy or risk of greater toxicity from targeted therapy.

Tumor progression, limited efficacy of current standard treatments, and the rise in patient mortality are associated with gene expression caused by the synergistic action of intratumoral hypoxia and HIF-1α activation. For this reason, recent investigations have focused on HIF-targeting therapeutic agents, with encouraging preclinical and clinical results in solid tumors. Here we describe the efficacy of a HIF-1α inhibitor, Acriflavine, and demonstrate its potency against brain cancer. This safe antibacterial dye induces cell death and apoptosis in several glioma cell lines, targets HIF-1α–mediated pathways, and decreases the level of PGK1, VEGF and HIF-1α in vitro and in vivo. Administered locally via biodegradable polymers, Acriflavine provides significant benefits in survival resulting in nearly 100% long term survival, confirmed by MRI and histological analyses. This study reports preclinical evidence that this safe, small molecule can contribute to brain tumor therapy and highlights the significance of HIF-1α-targeting molecules.

This is the peer reviewed version of the following article: “Puiggalí, A., Del Valle, L.J., Aleman, C. and Pérez, M. (2017) Weighing biointeractions between fibrin (ogen) and clot-binding peptides using microcantilever sensors. Journal of peptide science, (23) 2: 162–171.” which has been published in final form at [doi: 10.1002/psc.2938]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving." Peptides homing tumor vasculature are considered promising molecular imaging agents for cancer detection at an early stage. In addition to their high binding affinity, improved tissue penetrating ability, and low immunogenicity, they can deliver targeted anticancer drugs, thus expanding therapeutic treatments. Among those, CREKA, a linear peptide that specifically binds to clotted-plasma proteins in tumor vessels, has been recently employed to design bioactive systems able to target different cancer types. Within this context, this paper explores the biorecognition event between CR(NMe)EKA, an engineered CREKA-analog bearing a noncoded amino acid (N-methyl-Glu) that is responsible for its enhanced activity, and clotted-plasma proteins (fibrin and fibrinogen) by nanomechanical detection. Specifically, the tumor-homing peptide was covalently attached via epoxysilane chemistry onto silicon microcantilever chips that acted as sensors during dynamic mode experiments. Before that, each step of the functionalization process was followed by contact angle measurements, interferometry, X-ray photoelectron spectroscopy, and atomic force microscopy, thus revealing the applied protocol as a suitable strategy. The fibrin(ogen)-binding induced by CR(NMe)EKA was detected by the resonance frequency shift of the cantilevers, and a detection limit of 100¿ng/mL was achieved for both proteins. Even though further development is required, this work reflects the promising application of emerging technologies capable of assisting in the comprehension of biological interactions and their implications in the biotechnological field.