Search for new therapeutic targets for lung cancer

En cáncer de pulmón, una de las terapias dirigidas más exitosas, son las que inhiben a los protooncogenes ErbB (especialmente a EGFR y a ErbB2). Actualmente se están ensayando la inhibición de estos oncogenes mediante pequeñas moléculas (erlotinib, lapatinib, gefitinib) o mediante anticuerpos monoclonales (cetuximab, trastuzumab, pertuzumab) consiguiéndose un éxito relativo para tumores metastásicos de célula no pequeña. Interesantemente estas terapias parecen ser más efectivas cuando los oncogenes se encuentran amplificados. Teniendo en cuenta estas consideraciones, el objetivo principal de este trabajo, es la búsqueda de regiones cromosómicas amplificadas en cáncer de pulmón que puedan albergar nuevos oncogenes, que nos ayuden a profundizar en el conocimiento del cáncer y sobre los que podamos desarrollar futuros fármacos que sirvan como terapias dirigidas. Para tal propósito hemos combinado dos tipos de plataformas de análisis masivo de información genética: microarrays de hibridación genómica comparada (CGH) y microarrays de expresión para analizar el genoma de 8 líneas celulares de cáncer de pulmón. Se comparó directamente el número de copias de ADN y los niveles de expresión de ARNm de más de 8000 genes para identificar nuevos oncogenes amplificados funcionalmente. La línea celular Calu3 que contiene una amplificación cromosómica conocida de ErbB2, en la región 17q12, se utilizó como control positivo. Además de confirmar la amplificación en 17q en las células Calu3, nuestros datos revelaron claramente la presencia de cuatro amplicones independientes en los cromosomas 5p13, 6p21, 11q13 , 19q13 y en la mayoría de los casos de forma concomitante con un aumento en la expresión génica. Ensayos de fluorescencia in situ (FISH) en las líneas celulares confirmaron la presencia de amplificación génica (más de 10 copias) en estas regiones cromosómicas. Además de las líneas celulares, la presencia y la frecuencia de las alteraciones se evaluó adicionalmente en 26 tumores primarios de pulmón mediante FISH. Nuestros resultados mostraron una recurrencia en la amplificación cromosómica en tumores primarios: 8 % (en la región cromosómica 5p12), 4 % (6p21 ), 4 % (11q13.2 ), 2 % (17q12 ) y 2 % (19q13). Un análisis más detallado de los genes contenidos en estas regiones revelaron posibles candidatos a ser nuevos oncogenes en cáncer de pulmón. En conjunto, estos los datos sugieren que estas regiones cromosómicas de ampliación pueden albergar posibles oncogenes implicados en la carcinogénesis pulmonar sobre los cuales se podría diseñar de nuevos fármacos de utilidad terapéutica.In lung cancer, one of the most successful targeted therapies are those that inhibit ErbB protooncogenes (especially EGFR and ErbB2). Are currently being tested inhibition of these oncogenes by small molecules (erlotinib, lapatinib, gefitinib) or monoclonal antibodies (cetuximab, trastuzumab, pertuzumab) achieving relative success for metastatic tumors of non- small cell. Interestingly, these therapies are most effective when these oncogenes are amplified. Given these considerations, the main aim of this work is the search for chromosomal regions amplified in lung cancer that can accommodate new oncogenes, to help us deepen knowledge about cancer and future drugs that we develop to serve as targeted therapies. To search for chromosomal regions that could harbour new oncogenes amplified in lung tumors we have performed high-resolution CGH (Comparative Genome Hybridization), analysis on cDNA microarrays in 8 lung cancer cell lines. We directly compared DNA copy number and mRNA expression levels of over 8000 genes to identify novel amplicons and the candidate oncogenes that are targeted. The Calu-3 cells that carry gene amplification at the 17q12 region, a previously reported and well known amplicon that targets the ErbB2 oncogene, was included in the study as a positive control. In addition of confirming the amplification at 17q in the Calu-3 cells, our data clearly unveiled the presence of four independent amplicons at chromosomes 5p13, 6p21, 11q13, and 19q13, in most cases concomitantly with an increase in gene expression. FISH analysis in the cell lines confirmed the presence of gene amplification (more than 10 copies) at these regions. In addition to cell lines, the presence and frequency of the alterations was further evaluated in 26 lung primary tumors by FISH analysis. Our results showed that the frequency of amplification at these regions in the primary tumors was as follows: 8% (at chromosome 5p12), 4% (at chromosome 6p21), 4%(at chromosome 11q13.2), 2% (at chromosome 17q12) y 2% (at chromosome 19q13). Further analysis of the genes in these regions revealed potential candidates for new oncogenes in lung cancer. Taken together, the data suggests that these amplicons may harbour potential oncogenes implicated in lung carcinogénesis that could be used for drug targeting. Overall, our results illustrate how the amplification provides a powerful approach to highlight genes with an important role in cancer

Density-Temperature-Softness Scaling of the Dynamics of Glass-forming Soft-sphere Liquids

The principle of dynamic equivalence between soft-sphere and hard-sphere fluids [Phys. Rev. E \textbf{68}, 011405 (2003)] is employed to describe the interplay of the effects of varying the density n, the temperature T, and the softness (characterized by a softness parameter {\nu}^{-1}) on the dynamics of glass-forming soft-sphere liquids in terms of simple scaling rules. The main prediction is that the dynamic parameters of these systems, such as the {\alpha}-relaxation time and the long-time self-diffusion coefficient, depend on n, T, and {\nu} only through the reduced density n^\ast \equiv n{\sigma}^{3}_{HS}(T, {\nu}),where the effective hard-sphere diameter {\sigma}_{HS}(T, {\nu}) is determined, for example, by the Andersen-Weeks-Chandler condition for soft-sphere-hard-sphere structural equivalence. A number of scaling properties observed in recent simulations involving glass-forming fluids with repulsive short range interactions are found to be a direct manifestation of this general dynamic equivalence principle. The self-consistent generalized Langevin equation (SCGLE) theory of colloid dynamics is shown to accurately capture these scaling rule

Wiping DNA Methylation: Wip1 Regulates Genomic Fluidity on Cancer

Wip1 phosphatase plays an important role in cancer by inactivating p53 and INK4a/ARF pathways. In this issue of Cancer Cell, Filipponi and colleagues further connect the oncogenic role of Wip1 with heterochromatin dynamics, transposable element expression, and a mutation-prone environment that may enhance heterogeneity and ultimately contribute to tumor evolution

The dialogical potential of Facebook: the case of fashion brands

The main goal of this study is to examine the communication that fashion brands make through Facebook. It is intended to ascertain if the fashion brands use a communication strategy based on dialogue with their followers. In order to reach the objectives indicated above, a content analysis has been carried out (quantitative methodology) of 326 publications made through Facebook by H&M, Zara and Ralph Lauren between January 1st and June 30th of 2017. The main results of this work show that, although Facebook has a wide dialogical potential, fashion brands do not use it to a great extent. Although we consider appropriate to have analyzed only the social network Facebook given its im portance, this situation could be considered as the main limitation of the present study, since the fashion brands could, or not, be taking advantage of the dialogical potential of other social networks, such as Twitter or Instagram. It could be con sidered that this research stands out for studying a series of variables that allows organizations to know if they are using the dialogical potential offered by digital communication.info:eu-repo/semantics/publishedVersio

General Non-equilibrium Theory of Colloid Dynamics

A non-equilibrium extension of Onsager's canonical theory of thermal fluctuations is employed to derive a self-consistent theory for the description of the statistical properties of the instantaneous local concentration profile n(r,t) of a colloidal liquid in terms of the coupled time evolution equations of its mean value n(r,t) and of the covariance {\sigma}(r,r';t) \equiv of its fluctuations {\delta}n(r, t) = n(r, t) - n(r, t). These two coarse-grained equations involve a local mobility function b(r, t) which, in its turn, is written in terms of the memory function of the two-time correlation function C(r, r' ; t, t') \equiv <{\delta}n(r, t){\delta}n(r',t')>. For given effective interactions between colloidal particles and applied external fields, the resulting self-consistent theory is aimed at describing the evolution of a strongly correlated colloidal liquid from an initial state with arbitrary mean and covariance n^0(r) and {\sigma}^0(r,r') towards its equilibrium state characterized by the equilibrium local concentration profile n^(eq)(r) and equilibrium covariance {\sigma}^(eq)(r,r'). This theory also provides a general theoretical framework to describe irreversible processes associated with dynamic arrest transitions, such as aging, and the effects of spatial heterogeneities

Representation of defense organizations in the Marvel Cinematic Universe (2008-2019)

Homeland security and defense are common themes in superhero films. The Marvel Cinematic Universe (MCU) is formed by more than twenty films made over the course of a decade (2008-2019), during the government of three different Presidents of the United States. In this paper we analyze the diminishing representation of real defense forces in the MCU films. How the interference of the superhero is used as a narrative excuse to suggest a lack of or deficiencies in the law concerning freedom and defense is also studied, as well as how the films make a metaphorical discourse about the historical reality after September 11th

Novel and natural knockout lung cancer cell lines for the LKB1/STK11 tumor suppressor gene

Germline mutations of the LKB1 gene are responsible for Peutz-Jeghers syndrome (PJS), an autosomal dominant inherited disorder bestowing an increased risk of cancer. We have recently demonstrated that LKB1 inactivating mutations are not confined to PJS, but also appear in lung adenocarcinomas of sporadic origin, including primary tumors and lung cancer cell lines. To accurately determine the frequency of inactivating LKB1 gene mutations in lung tumors we have sequenced the complete coding region of LKB1 in 21 additional lung cancer cell lines. Here we describe the mutational status of LKB1 gene in 30 lung cancer cell lines from different histopathological types, including 11 lung adenocarcinomas (LADs) and 11 small cell lung cancers (SCLCs). LKB1 gene alterations were present in six (54%) of the LAD cell lines tested but in none of the other histological types. Similar to our previous observations in primary tumors, all point mutations were of the nonsense or frameshift type, leading to an abnormal, truncated protein. Moreover, 2 cell lines (A427 and H2126) harbored large gene deletions that spanned several exons. Hence, we have identified additional lung cancer cell lines carrying inactivating mutations of the LKB1 tumor suppressor gene, further attesting to the significance of this gene in the development of LADs and providing new natural LKB1 knockouts for studies of the biological function of the LKB1 protein

The African hind's (Cephalopholis taeniops, serranidae) use of artificial reefs off Sal Island (Cape Verde): a preliminary study based on acoustic telemetry

The African hind Cephalopholis taeniops (Valenciennes, 1828) is one of the most important commercial demersal species caught in the Cape Verde archipelago. The species is closely associated with hard substrate and is one of the main attractions for SCUBA divers. In January 2006 a former Soviet fishing vessel - the Kwarcit - was sunk off Santa Maria Bay (Sal Island). Young C. taeniops are commonly observed in this artificial reef (AR). In order to investigate the species' use of the AR, 4 specimens were captured and surgically implanted underwater with Vemco brand acoustic transmitters. The fish were monitored daily with an active telemetry receiver for one week after release. Simultaneously, an array of 3 passive VR2 / VR2W receivers was set for 63 days, registering data that allowed an analysis of spatial, daily and short term temporal activity patterns. The results showed site fidelity to the AR, with no migrations to the nearby natural reef. The method used allowed to register a consistent higher activity during daytime and a preference for the area opposite the dominant current

Genome-wide CRISPR interference screen identifies long non-coding RNA loci required for differentiation and pluripotency

Although many long non-coding RNAs (lncRNAs) exhibit lineage-specific expression, the vast majority remain functionally uncharacterized in the context of development. Here, we report the first described human embryonic stem cell (hESC) lines to repress (CRISPRi) or activate (CRISPRa) transcription during differentiation into all three germ layers, facilitating the modulation of lncRNA expression during early development. We performed an unbiased, genome-wide CRISPRi screen targeting thousands of lncRNA loci expressed during endoderm differentiation. While dozens of lncRNA loci were required for proper differentiation, most differentially expressed lncRNAs were not, supporting the necessity for functional screening instead of relying solely on gene expression analyses. In parallel, we developed a clustering approach to infer mechanisms of action of lncRNA hits based on a variety of genomic features. We subsequently identified and validated FOXD3-AS1 as a functional lncRNA essential for pluripotency and differentiation. Taken together, the cell lines and methodology described herein can be adapted to discover and characterize novel regulators of differentiation into any lineage

Metabolites of the gut microbiota involved in the autism spectrum disorder

En los últimos años, ha habido un aumento de los estudios que buscan comprender la relación existente entre el microbiota intestinal (MI) con el trastorno del espectro autista (TEA), que debe producirse a través del eje microbiota-intestino-cerebro. A pesar de que los distintos autores señalan que los cambios encontrados en distintos filos, familias y géneros bacterianos están implicados en el TEA, no hay consenso científico a día de hoy. Algunos autores apuntan a la posible relación existente entre dichas poblaciones bacterianas con ciertos productos de excreción o metabolitos como el ácido propiónico ya que aparecen con frecuencia en niños con TEA. Aunque en los últimos años la MI comienza a acumular evidencia científica, en términos de neurociencia, el estudio de la metabolómica asociada a la misma y los mecanismos mediante los cuales estos metabolitos pueden influir en la aparición y desarrollo del TEA aún permanece en sus primeros estadios.In recent years, there has been an increase in studies that seek to understand the relationship between gut microbiota (GM) with the behavior of people with autism spectrum disorders (ASD), which must occur through the microbiota-gut-brain axis. Although the different authors point out that the changes found in different phyla, families and bacterial genera are involved in ASD, there is no scientific consensus to date. Some authors point to the possible relationship between these bacterial populations with certain products of excretion or metabolites such as propionic acid since they frequently appear in children with ASD. Although in recent years the GM has begun to accumulate scientific evidence, in terms of neuroscience, the study of the metabolomics associated with it and the mechanisms by which these metabolites can influence the appearance and development of ASD remains in its first stages