Identification and characterization of cancer initiating cells in K-Ras driven Non Small Cell Lung Cancer

Tesis doctoral inédita. Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 10-02-2013Hemos estudiado los primeros eventos que acompañan la transformación inducida por el oncogen K-­‐Ras en el modelo de adenocarcinoma pulmonar K-­‐RasLSLG12Vgeo. Por un lado, hemos seguido la expansión de los distintos tipos celulares del pulmón tras la activación del oncogen K-­‐Ras con el fin de identificar las células iniciadoras del cancer de pulmón. Hemos descubierto que las células alveolares de tipo II son las que con más probabilidad dan origen a los adenocarcinomas de pulmón cuando la expresión del oncogen se produce de forma aleatoria. Sin embargo, la activación preferential de K-­‐ RasG12V en el epitelio bronquiolar a través de la infección intratraqueal con el virus adeno-­‐Cre, así como la expresión del oncogen durante el desarrollo embrionario del pulmón, producen una hiperproliferación de las células Clara. Por otro lado, hemos obtenido el perfil de expresión genica de la población celular que se expande tras la expresión de K-­‐RasG12V a tiempos muy tempranos. Así, hemos podido identificar el factor de transcripción de la familia Ets, Etv4, como un mediador muy importante de la transformación inducida por K-­‐RasG12V.We have investigated the first steps of K-­‐RasG12V-­‐induced transformation in the K-­‐ RasLSLG12Vgeo model of lung adenocarcinoma. One one side, we have followed expansion of the different lung cell types after K-­‐RasG12V –activation in order to identify lung cancer initiating cells. We have found that alveolar type II cells are the most likely to initiate lung cancer when the oncogene activation occurrs in an unbiased fashion. On the contrary, preferential expression of K-­‐RasG12V in the bronchiolar epithelium via adeno-­‐ Cre intratracheal infection or activation of the oncogene during lung embryonic development, leads to hyperproliferation of Clara cells. On the other side, we obtained the gene expression profile of the cells that expand following oncogenic K-­‐Ras activation at very early time points. Thus, we have identified the Ets transcription factor Etv4 as a key mediator of K-­‐RasG12V-­‐induced transformation

Non-invasive evaluation of the effect of metoprolol on the atrioventricular node during permanent atrial fibrillation.

During atrial fibrillation (AF), conventional electrophysiological techniques for assessment of refractory period or conduction velocity of the atrioventricular (AV) node cannot be used. We aimed at evaluating changes in AV nodal properties during administration of metoprolol from electrocardiogram data, and to support our findings with simulated data based on results from an electrophysiological study

Avaliação da atividade antibacteriana in vitro do medicamento ácido acetilsalicílico

Objetivo: Avaliar a atividade antibacteriana do anti-inflamatório ácido acetilsalicílico (AAS) frente seis cepas bacterianas padrão de referência American Type Culture Collection (ATCC) e 41 isolados clínicos multidroga resistentes (MDR). Métodos: A atividade antibacteriana do medicamento foi determinada através da concentração inibitória mínima (CIM) obtida frente às cepas bacterianas. Resultados: AAS demonstrou atividade antibacteriana tanto frente a cepas Gram negativas (GN) como Gram positivas (GP), apresentando, frente a maioria das cepas testadas, CIM de 1024 µg/mL e 2048 µg/mL. Frente a um isolado clínico de Acinetobacter baumannii MDR, apresentou CIM de 512 µg/mL. AAS demonstrou atividade antibacteriana frente a microrganismos altamente resistentes. Conclusão: Nossos resultados permitem inferir que o medicamento AAS apresenta potencial atividade antibacteriana frente às cepas MDR testadas, porém estudos complementares devem ser realizados a fim de identificar seu mecanismo de ação na inibição bacteriana. Testes de sinergismos também são aconselhados com o objetivo de avaliar a associação do AAS com antibacterianos já utilizados na clínica, visto que sua atividade pode ser potencializada

Combining robotics with enhanced serotonin-driven cortical plasticity improves post-stroke motor recovery.

Despite recent progresses in robotic rehabilitation technologies, their efficacy for post-stroke motor recovery is still limited. Such limitations might stem from the insufficient enhancement of plasticity mechanisms, crucial for functional recovery. Here, we designed a clinically relevant strategy that combines robotic rehabilitation with chemogenetic stimulation of serotonin release to boost plasticity. These two approaches acted synergistically to enhance post-stroke motor performance. Indeed, mice treated with our combined therapy showed substantial functional gains that persisted beyond the treatment period and generalized to non-trained tasks. Motor recovery was associated with a reduction in electrophysiological and neuroanatomical markers of GABAergic neurotransmission, suggesting disinhibition in perilesional areas. To unveil the translational potentialities of our approach, we specifically targeted the serotonin 1A receptor by delivering Buspirone, a clinically approved drug, in stroke mice undergoing robotic rehabilitation. Administration of Buspirone restored motor impairments similarly to what observed with chemogenetic stimulation, showing the immediate translational potential of this combined approach to significantly improve motor recovery after stroke

O espaço da mulher na sociedade: uma reflexão a partir de o Segundo Sexo de Simone de Beauvoir

Este artigo busca analisar, através da obra de Simone de Beauvoir: O segundo sexo, o espaço da mulher na sociedade. A obra foi estudada na disciplina de Filosofia Contemporânea do curso de Licenciatura em Filosofia pela Universidade do Estado de Mato Grosso – UNEMAT. Tem por objetivo expor as dificuldades enfrentadas pelas mulheres no decorrer da história até a contemporaneidade, com a intenção de destacar a luta e sua coragem frente a uma sociedade construída pelos homens e para os homens, destacando o nascimento de muitos direitos conquistados pelo sexo feminino. Sendo assim, a pesquisa aborda o movimento feminista e a sua importância diante de todo processo de busca por respeito pelo segundo sexo, expondo um olhar sobre o que significava ser mulher, sob a ótica de como a mulher era vista e o que esperavam dela, comparando-se com os dias atuais. Usando Simone de Beauvoir como referência de personalidade feminina, como alguém que alcançou um espaço valioso para todas as mulheres.

Extensive preclinical validation of combined RMC-4550 and LY3214996 supports clinical investigation for KRAS mutant pancreatic cancer

Over 90% of pancreatic cancers present mutations in KRAS, one of the most common oncogenic drivers overall. Currently, most KRAS mutant isoforms cannot be targeted directly. Moreover, targeting single RAS downstream effectors induces adaptive resistance mechanisms. We report here on the combined inhibition of SHP2, upstream of KRAS, using the allosteric inhibitor RMC-4550 and of ERK, downstream of KRAS, using LY3214996. This combination shows synergistic anti-cancer activity in vitro, superior disruption of the MAPK pathway, and increased apoptosis induction compared with single-agent treatments. In vivo, we demonstrate good tolerability and efficacy of the combination, with significant tumor regression in multiple pancreatic ductal adenocarcinoma (PDAC) mouse models. Finally, we show evidence that 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) can be used to assess early drug responses in animal models. Based on these results, we will investigate this drug combination in the SHP2 and ERK inhibition in pancreatic cancer (SHERPA; ClinicalTrials.gov: NCT04916236) clinical trial, enrolling patients with KRAS-mutant PDAC.This work was funded by the American Association for Cancer Research, Lustgarten Foundation, and Stand Up to Cancer as a Pancreatic Cancer Collective New Therapies Challenge grant (grant no. SU2C-AACR-PCC-01-18)

A large-scale organoid-based screening platform to advance drug repurposing in pancreatic cancer

Hirt et al.1 report an automated, high-throughput drug screening platform for organoid cultures to enable repurposing of previously approved drugs for pancreatic cancers harboring specific genetic alterations. The pancreatic cancer organoid biobank also represents a valuable tool to uncover new drug-gene interactions in pancreatic tumors

Identification of cancer initiating cells in K-Ras driven lung adenocarcinoma

Ubiquitous expression of a resident K-RasG12V oncogene in adult mice revealed that most tissues are resistant to K-Ras oncogenic signals. Indeed, K-RasG12V expression only induced overt tumors in lungs. To identify these transformation-permissive cells, we induced K-RasG12V expression in a very limited number of adult lung cells (0.2%) and monitored their fate by X-Gal staining, a surrogate marker coexpressed with the K-Ras G12V oncoprotein. Four weeks later, 30% of these cells had proliferated to form small clusters. However, only SPC+ alveolar type II (ATII) cells were able to form hyperplastic lesions, some of which progressed to adenomas and adenocarcinomas. In contrast, induction of K-Ras G12V expression in lung cells by intratracheal infection with adenoviral-Cre particles generated hyperplasias in all regions except the proximal airways. Bronchiolar and bronchioalveolar duct junction hyperplasias were primarily made of CC10+ Clara cells. Some of them progressed to form benign adenomas. However, only alveolar hyperplasias, exclusively made up of SPC+ ATII cells, progressed to yield malignant adenocarcinomas. Adenoviral infection induced inflammatory infiltrates primarily made of T and B cells. This inflammatory response was essential for the development of K-RasG12V-driven bronchiolar hyperplasias and adenomas, but not for the generation of SPC+ ATII lesions. Finally, activation of K-RasG12V during embryonic development under the control of a Sca1 promoter yielded CC10+, but not SPC+, hyperplasias, and adenomas. These results, taken together, illustrate that different types of lung cells can generate benign lesions in response to K-Ras oncogenic signals. However, in adult mice, only SPC+ ATII cells were able to yield malignant adenocarcinomas.Work was supported by European Research Council Grant ERC-AG/250297-RAS AHEAD; EU-Framework Programme Grants LSHG-CT-2007-037665/CHEMORES, HEALTH-F2-2010-259770/LUNGTARGET, and HEALTH-010-260791/EUROCANPLATFORM; Spanish Ministry of Economy and Competitiveness Grant SAF2011-30173; Autonomous Community of Madrid S2011/BDM-2470/ONCOCYCLE (to M.B.); National Institutes of Health Grant R01 CA109335-04A1; Spanish Ministry of Economy and Competitivenes Grant SAF2012-32810; and EU-Framework Programme Grant FP7-ENV-2011/ARIMMORA (to I.S.-G.). S.M. was supported by a predoctoral fellowship from Fundación La Caixa.Peer Reviewe