Discovery of senolytics using machine learning.

Cellular senescence is a stress response involved in ageing and diverse disease processes including cancer, type-2 diabetes, osteoarthritis and viral infection. Despite growing interest in targeted elimination of senescent cells, only few senolytics are known due to the lack of well-characterised molecular targets. Here, we report the discovery of three senolytics using cost-effective machine learning algorithms trained solely on published data. We computationally screened various chemical libraries and validated the senolytic action of ginkgetin, periplocin and oleandrin in human cell lines under various modalities of senescence. The compounds have potency comparable to known senolytics, and we show that oleandrin has improved potency over its target as compared to best-in-class alternatives. Our approach led to several hundred-fold reduction in drug screening costs and demonstrates that artificial intelligence can take maximum advantage of small and heterogeneous drug screening data, paving the way for new open science approaches to early-stage drug discovery.

Reversal of mitochondrial malate dehydrogenase 2 enables anaplerosis via redox rescue in respiration-deficient cells.

Inhibition of the electron transport chain (ETC) prevents the regeneration of mitochondrial NAD+, resulting in cessation of the oxidative tricarboxylic acid (TCA) cycle and a consequent dependence upon reductive carboxylation for aspartate synthesis. NAD+ regeneration alone in the cytosol can rescue the viability of ETC-deficient cells. Yet, how this occurs and whether transfer of oxidative equivalents to the mitochondrion is required remain unknown. Here, we show that inhibition of the ETC drives reversal of the mitochondrial aspartate transaminase (GOT2) as well as malate and succinate dehydrogenases (MDH2 and SDH) to transfer oxidative NAD+ equivalents into the mitochondrion. This supports the NAD+-dependent activity of the mitochondrial glutamate dehydrogenase (GDH) and thereby enables anaplerosis-the entry of glutamine-derived carbon into the TCA cycle and connected biosynthetic pathways. Thus, under impaired ETC function, the cytosolic redox state is communicated into the mitochondrion and acts as a rheostat to support GDH activity and cell viability.

Toll-like receptor 2 orchestrates a tumor suppressor response in non-small cell lung cancer.

Targeting early-stage lung cancer is vital to improve survival. However, the mechanisms and components of the early tumor suppressor response in lung cancer are not well understood. In this report, we study the role of Toll-like receptor 2 (TLR2), a regulator of oncogene-induced senescence, which is a key tumor suppressor response in premalignancy. Using human lung cancer samples and genetically engineered mouse models, we show that TLR2 is active early in lung tumorigenesis, where it correlates with improved survival and clinical regression. Mechanistically, TLR2 impairs early lung cancer progression via activation of cell intrinsic cell cycle arrest pathways and the proinflammatory senescence-associated secretory phenotype (SASP). The SASP regulates non-cell autonomous anti-tumor responses, such as immune surveillance of premalignant cells, and we observe impaired myeloid cell recruitment to lung tumors after Tlr2 loss. Last, we show that administration of a TLR2 agonist reduces lung tumor growth, highlighting TLR2 as a possible therapeutic target.

MYC sensitises cells to apoptosis by driving energetic demand.

The MYC oncogene is a potent driver of growth and proliferation but also sensitises cells to apoptosis, which limits its oncogenic potential. MYC induces several biosynthetic programmes and primary cells overexpressing MYC are highly sensitive to glutamine withdrawal suggesting that MYC-induced sensitisation to apoptosis may be due to imbalance of metabolic/energetic supply and demand. Here we show that MYC elevates global transcription and translation, even in the absence of glutamine, revealing metabolic demand without corresponding supply. Glutamine withdrawal from MRC-5 fibroblasts depletes key tricarboxylic acid (TCA) cycle metabolites and, in combination with MYC activation, leads to AMP accumulation and nucleotide catabolism indicative of energetic stress. Further analyses reveal that glutamine supports viability through TCA cycle energetics rather than asparagine biosynthesis and that TCA cycle inhibition confers tumour suppression on MYC-driven lymphoma in vivo. In summary, glutamine supports the viability of MYC-overexpressing cells through an energetic rather than a biosynthetic mechanism.

In Vivo Modeling of Patient Genetic Heterogeneity Identifies New Ways to Target Cholangiocarcinoma.

Intrahepatic cholangiocarcinoma (ICC) is an aggressive malignancy of the bile ducts within the liver characterized by high levels of genetic heterogeneity. In the context of such genetic variability, determining which oncogenic mutations drive ICC growth has been difficult, and developing modes of patient stratification and targeted therapies remains challenging. Here we model the interactions between rare mutations with more common driver genes and combine in silico analysis of patient data with highly multiplexed in vivo CRISPR-spCas9 screens to perform a functional in vivo study into the role genetic heterogeneity plays in driving ICC. Novel tumor suppressors were uncovered, which, when lost, cooperate with the RAS oncoprotein to drive ICC growth. Focusing on a set of driver mutations that interact with KRAS to initiate aggressive, sarcomatoid-type ICC revealed that tumor growth relies on Wnt and PI3K signaling. Pharmacologic coinhibition of Wnt and PI3K in vivo impeded ICC growth regardless of mutational profile. Therefore, Wnt and PI3K activity should be considered as a signature by which patients can be stratified for treatment independent of tumor genotype, and inhibitors of these pathways should be levied to treat ICC. SIGNIFICANCE: This work shows that, despite significant genetic heterogeneity, intrahepatic cholangiocarcinoma relies on a limited number of signaling pathways to grow, suggesting common therapeutic vulnerabilities across patients.

Cytoplasmic innate immune sensing by the caspase-4 non-canonical inflammasome promotes cellular senescence.

Cytoplasmic recognition of microbial lipopolysaccharides (LPS) in human cells is elicited by the caspase-4 and caspase-5 noncanonical inflammasomes, which induce a form of inflammatory cell death termed pyroptosis. Here we show that LPS-mediated activation of caspase-4 also induces a stress response promoting cellular senescence, which is dependent on the caspase-4 substrate gasdermin-D and the tumor suppressor p53. Furthermore, we found that the caspase-4 noncanonical inflammasome is induced and assembled in response to oncogenic RAS signaling during oncogene-induced senescence (OIS). Moreover, targeting caspase-4 expression in OIS showed its critical role in the senescence-associated secretory phenotype and the cell cycle arrest induced in cellular senescence. Finally, we observed that caspase-4 induction occurs in vivo in mouse models of tumor suppression and ageing. Altogether, we are showing that cellular senescence is induced by cytoplasmic LPS recognition by the noncanonical inflammasome and that this pathway is conserved in the cellular response to oncogenic stress.

Termorregulación de la lagartija Liolaemus chacoensis (Squamata: Liolaemidae) durante su ciclo reproductivo, en el Chaco occidental, Argentina

La regulación de la temperatura corporal en las lagartijas juega un rol clave para la supervivencia de las especies. Su condición fisiológica, en referencia a su dependencia de la temperatura ambiente, los convierte en un excelente modelo de estudio. Asimismo, los cambios en el ciclo reproductivo provocan modificaciones en los parámetros termofisiológicos. Este estudio es el primero en abordar la relación de la termorregulación con la actividad reproductiva y el sexo, para una especie de Liolaemus de la ecorregión del Chaco. El objetivo de este trabajo fue determinar cómo influye el período reproductivo, de reclutamiento y el pos-reproductivo, así como también el efecto del sexo y la longitud-hocico-cloaca sobre la termorregulación de Liolaemus chacoensis. Los individuos fueron capturados a mano en el sector Sur de Valle Fértil, provincia de San Juan, Argentina. Se capturaron un total de 44 individuos durante noviembre 2014, febrero 2015 y marzo 2015. Se registró la temperatura corporal (Tc), la temperatura del sustrato (Ts) y del aire (Ta). También, se utilizaron modelos biofísicos para registrar la temperatura operativa. Posteriormente, en el laboratorio se evaluó la temperatura seleccionada (Tsel). Luego se calculó el índice de eficiencia termorregulatoria. Los resultados indicaron que la Tc de L. chacoensis se relacionó positivamente con la Ta, presentando un comportamiento heliotérmico que se mantuvo durante los períodos y el sexo. Asimismo, la Tc fue mayor para el período reproductivo y de reclutamiento con respecto al pos-reproductivo. En particular, solo el sexo tuvo efecto sobre la Tsel, sin embargo para el rango intercuartil de Tsel tuvo efecto tanto el período del ciclo reproductivo como el sexo. El índice de eficiencia termorregulatoria indicó que L. chacoensis es un termoconforme absoluto. Sin embargo, en el período reproductivo tanto machos como hembras cambian hacia la termorregulación activa. Estos resultados muestran que los parámetros termofisiológicos, varían en los diferentes períodos del ciclo reproductivo y la selección activa de micrositios sería un mecanismo para el mantenimiento de la temperatura corporal de la especie.

Thermoregulation of Liolaemus chacoensis lizard (Squamata: Liolaemidae) during its reproductive cycle, in the Western Chaco, Argentina. Regulation of body temperature in lizards plays a key role in the survival of species. Their physiological condition, in reference to their dependence on the ambient temperature makes them an excellent study model. Also, changes in the reproductive cycle cause changes in thermophysiological parameters. This study is the first to address the relationship of thermoregulation with reproductive activity and sex, for a species of Liolaemus from the Chaco ecoregion. The aim of this work was to determine how the reproductive, recruitment and post-reproductive periods influences, as well as the effect of sex and snout-vent length on the thermoregulation of Liolaemus chacoensis. Individuals were captured by hand in the Southern sector of Valle Fertil, San Juan province, Argentina. A total of 44 individuals were captured during the months of November 2014, February 2015 and March 2015. Body (Tb), substrate (Ts) and air (Ta) temperatures were recorded. Also, biophysical models were used to record operative temperatures. Later selected temperatures (Tsel) were evaluated at the laboratory. Finally, thermoregulatory efficiency index was calculated. Results indicated that L. chacoensis Tb was positively related to Ta, presenting a heliothermic behavior that was maintained during periods and sex. Also, Tb was higher in reproductive and recruitment periods than in post-reproductive period. In particular, only sex had an effect on Tsel, however for the interquartile range of Tsel both the period of reproductive cycle and sex had an effect. The thermoregulatory efficiency index indicated that L. chacoensis is an absolute thermoconformist. However, in the reproductive period both males and females change towards active thermoregulation. These results show that thermophysiological parameters vary in different periods of reproductive cycle and the active microsite selection would be a mechanism in maintenance of body temperature of this species.

Inhibition of the 60S ribosome biogenesis GTPase LSG1 causes endoplasmic reticular disruption and cellular senescence.

Cellular senescence is triggered by diverse stimuli and is characterized by long-term growth arrest and secretion of cytokines and chemokines (termed the SASP-senescence-associated secretory phenotype). Senescence can be organismally beneficial as it can prevent the propagation of damaged or mutated clones and stimulate their clearance by immune cells. However, it has recently become clear that senescence also contributes to the pathophysiology of aging through the accumulation of damaged cells within tissues. Here, we describe that inhibition of the reaction catalysed by LSG1, a GTPase involved in the biogenesis of the 60S ribosomal subunit, leads to a robust induction of cellular senescence. Perhaps surprisingly, this was not due to ribosome depletion or translational insufficiency, but rather through perturbation of endoplasmic reticulum homeostasis and a dramatic upregulation of the cholesterol biosynthesis pathway. The underlying transcriptomic signature is shared with several other forms of senescence, and the cholesterol biosynthesis genes contribute to the cell cycle arrest in oncogene-induced senescence. Furthermore, targeting of LSG1 resulted in amplification of the cholesterol/ER signature and restoration of a robust cellular senescence response in transformed cells, suggesting potential therapeutic uses of LSG1 inhibition.

The innate immune sensor Toll-like receptor 2 controls the senescence-associated secretory phenotype.

Cellular senescence is a stress response program characterized by a robust cell cycle arrest and the induction of a proinflammatory senescence-associated secretory phenotype (SASP) that is triggered through an unknown mechanism. Here, we show that, during oncogene-induced senescence (OIS), the Toll-like receptor 2 (TLR2) and its partner TLR10 are key mediators of senescence in vitro and in murine models. TLR2 promotes cell cycle arrest by regulating the tumor suppressors p53-p21CIP1, p16INK4a, and p15INK4b and regulates the SASP through the induction of the acute-phase serum amyloids A1 and A2 (A-SAAs) that, in turn, function as the damage-associated molecular patterns (DAMPs) signaling through TLR2 in OIS. Last, we found evidence that the cGAS-STING cytosolic DNA sensing pathway primes TLR2 and A-SAAs expression in OIS. In summary, we report that innate immune sensing of senescence-associated DAMPs by TLR2 controls the SASP and reinforces the cell cycle arrest program in OIS.

Notch Signaling Mediates Secondary Senescence.

Oncogene-induced senescence (OIS) is a tumor suppressive response to oncogene activation that can be transmitted to neighboring cells through secreted factors of the senescence-associated secretory phenotype (SASP). Currently, primary and secondary senescent cells are not considered functionally distinct endpoints. Using single-cell analysis, we observed two distinct transcriptional endpoints, a primary endpoint marked by Ras and a secondary endpoint marked by Notch activation. We find that secondary oncogene-induced senescence in vitro and in vivo requires Notch, rather than SASP alone, as previously thought. Moreover, Notch signaling weakens, but does not abolish, SASP in secondary senescence. Global transcriptomic differences, a blunted SASP response, and the induction of fibrillar collagens in secondary senescence point toward a functional diversification between secondary and primary senescence.

Nuclear pore density controls heterochromatin reorganization during senescence.

During oncogene-induced senescence (OIS), heterochromatin is lost from the nuclear periphery and forms internal senescence-associated heterochromatin foci (SAHFs). We show that an increased nuclear pore density during OIS is responsible for SAHF formation. In particular, the nucleoporin TPR is necessary for both formation and maintenance of SAHFs. Loss of SAHFs does not affect cell cycle arrest but abrogates the senescence-associated secretory phenotype-a program of inflammatory cytokine gene activation. Our results uncover a previously unknown role of nuclear pores in heterochromatin reorganization in mammalian nuclei and demonstrate the importance of heterochromatin organization for a specific gene activation program.

Measuring the Inflammasome in Oncogene-Induced Senescence.

Inflammasomes are multimeric protein complexes that process IL-1ß by cleaving the translated full-length protein into its active IL-1ß mature fragment. In oncogene-induced senescence, inflammasomes play a crucial role by regulating IL1R signaling and consequently modulating proliferation and the senescence-associated secretory phenotype (SASP). Inflammasome activation requires two steps: (a) priming of the inflammasome by activation of IL1B expression, followed by (b) cleavage and release of mature IL-1ß. In this chapter, we describe methods to detect both stages of inflammasome activation in cellular senescence.

Comportamiento de las neumonías complicadas en niños en hospital pediátrico provincial pinareño / Behavior of complicated pneumonias in children at pediatric provincial hospital of Pinar del Rio

RESUMEN Introducción: las neumonías representan uno de los principales problemas de Salud Pública en menores de cinco años a nivel mundial, constituyen la primera causa de mortalidad por infecciones agudas y la segunda de hospitalización. Objetivo: caracterizar clínico y epidemiológicamente las neumonías complicadas en niños ingresados en el servicio de Enfermedades Respiratorias del Hospital Pediátrico Provincial "Pepe Portilla" durante el período 2015-2016. Métodos: se realizó una investigación observacional, descriptiva y transversal, el universo estuvo conformado por los pacientes entre un mes y 14 años ingresados por neumonía, y la muestra quedó conformada por 50 casos con evolución complicada. Se utilizaron las historias clínicas de los pacientes y un cuestionario aplicado a los padres. Los datos se mostraron empleando frecuencias absolutas y relativas porcentuales. Resultados: predominó el grupo de edades de 1-4 años (28 %), el sexo masculino (64 %) y los pertenecientes al área urbana (58 %). Las enfermedades respiratorias alérgicas constituyeron la principal enfermedad crónica asociada. El bajo peso al nacer, la desnutrición proteico energética, el abandono precoz de la lactancia materna, la convivencia con fumadores y la asistencia a círculos infantiles fueron factores de riesgo detectados en los pacientes estudiados. La complicación fundamental fue el derrame pleural (68 %). Conclusiones: se evidenció que la neumonía complicada en el período señalado es un problema de salud que orienta al diseño de acciones para prevenir la presentación de las complicaciones.

ABSTRACT Introduction: pneumonias represent one of the main public health problems worldwide in children under five years old; they constitute the first cause of mortality due to acute infections and the second cause of hospitalization. Objective: to characterize clinically and epidemiologically complicated pneumonias in children admitted Respiratory Diseases Service at Pepe Portilla Provincial Pediatric Hospital during 2015-2016. Methods: an observational, descriptive and cross-sectional research was carried out. The target group consisted of patients between one month and 14 years old hospitalized for pneumonia, and the sample consisted of 50 cases with complicated evolution. The patient's medical records and a questionnaire applied to the parents were analyzed; using absolute and relative percentage frequencies to express the data. Results: the age group of 1-4 years (28 %), male sex (64 %) and those belonging to the urban area (58 %) predominated. Allergic respiratory diseases were the main associated chronic disease. Low birth-weight, energy protein malnutrition, early abandonment of breastfeeding, cohabitation with smokers and attendance at children's day care centers were the risk factors detected in the patients studied. The main complication was pleural effusion (68 %). Conclusions: complicated pneumonia was characterized in the indicated period, detecting elements that show these are still a health problem and guide the design of actions to prevent the onset of complications.

Patrones de herbivoría en Vassobia breviflora (Solanaceae): variación en el daño foliar y selección natural mediada por herbívoros / Patterns of herbivory in Vassobia breviflora (Solanaceae): variation in foliar damage and natural selection mediated by herbivorous

Resumen La selección mediada por herbívoros moldea la evolución de los caracteres defensivos en las plantas. El conocimiento acerca del rol de los herbívoros como mediadores de selección es escaso y más aún si se consideran los grupos funcionales de herbívoros. Los objetivos de este trabajo fueron (1) describir la variación en rasgos foliares entre poblaciones, entre plantas dentro de una población y a nivel sub-individual (intra-planta), (2) explorar la relación entre la variación en el nivel de herbivoría y rasgos foliares, (3) determinar la relación entre rasgos foliares y patrones de daño y (4) estimar los regímenes de selección por diferentes grupos funcionales de herbívoros. Realizamos este estudio en cuatro poblaciones de Vassobia breviflora en el noroeste de Argentina (Yungas). Los rasgos foliares considerados fueron: tamaño, área foliar (af), forma (relación longitud/ancho de la hoja; laf) y proporción de área foliar removida (pafr) (N = 1 582 hojas, 57 plantas). Los herbívoros consumieron 15.6 % del área foliar y 76.8 % de la variación en la pafr ocurrió a nivel sub-individual. El patrón de daño fue dominado por herbívoros cortadores (70 %), seguido de un patrón de herbivoría punteada (22 %), 5 % mixto y 1 % minador. Se detectó selección no lineal para laf (γii = -0.180, EE = 0.76, P < 0.05), y selección correlacional entre el daño cortador y af (γij = -1.297, SE = 0.62, P < 0.05) y entre el daño punteado y af (γij= -1.130, SE=0.76, P < 0.05). La selección natural favoreció plantas con hojas pequeñas y alta remoción foliar y hojas grandes con menor daño y se detectó selección en contra de hojas grandes con mayor daño. Además, deducido de la relación entre el tipo de daño y la adecuación relativa, la selección favorecería el daño punteado por sobre el cortador. Las plantas resolverían el conflicto con los herbívoros según el tipo de daño y la selección natural regularía el despliegue foliar como una estrategia para lidiar con la diversidad de herbívoros.(AU)

Abstract Herbivore mediated-selection shapes the evolution of defensive plant traits. Knowledge about the role of herbivores as mediators of selection is scarce and even more if herbivore functional groups are considered. The objectives of this work were (1) to describe the variation in foliar traits between populations and both between and intra-plants within a population, (2) to explore the relationship between the variation in the herbivory level and foliar traits, (3) to determine the relationship between leaf traits and damage patterns and (4) estimate the selection regimes by different herbivore functional groups. We conducted this study in four populations of Vassobia breviflora in Northwestern Argentina (Yungas). The foliar traits considered were size, leaf area (af), shape (leaf length / width ratio; laf) and proportion of leaf area removed (pafr) (N = 1 582 leaves, 57 plants). The herbivores consumed 15.6 % of the leaf area and 76.8 % of the variation in the pafr occurred at the sub-individual level. The damage pattern was dominated by cutter herbivores (70 %), followed by a dotted herbivory pattern (22 %), mixed 5 % and 1 % miner. Nonlinear selection was detected for laf (γii = -0.180; EE = 0.76; P < 0.05), and correlational selection between the cutter damage and af (γij = -1.297; SE = 0.62; P < 0.05) and between the dotted damage and af (γij = -1.130; SE = 0.76; P < 0.05). Natural selection favored plants with small leaves and high foliar removal and large leaves with less damage and selection against larger leaves with greater damage was detected. In addition, deduced from the relationship between the damage type and the relative fitness, the selection would favor the dotted damage over the cutter one. The plants would resolve the conflict with the herbivores according to the damage type and natural selection would regulate the foliar display as a strategy to deal with the herbivore diversity.(AU)

PHD3 Regulates p53 Protein Stability by Hydroxylating Proline 359.

Cellular p53 protein levels are regulated by a ubiquitination/de-ubiquitination cycle that can target the protein for proteasomal destruction. The ubiquitination reaction is catalyzed by a multitude of ligases, whereas the removal of ubiquitin chains is mediated by two deubiquitinating enzymes (DUBs), USP7 (HAUSP) and USP10. Here, we show that PHD3 hydroxylates p53 at proline 359, a residue that is in the p53-DUB binding domain. Hydroxylation of p53 upon proline 359 regulates its interaction with USP7 and USP10, and its inhibition decreases the association of p53 with USP7/USP10, increases p53 ubiquitination, and rapidly reduces p53 protein levels independently of mRNA expression. Our results show that p53 is a PHD3 substrate and that hydroxylation by PHD3 regulates p53 protein stability through modulation of ubiquitination.

Purulent pericarditis as a complication of pneumonia in an infant. clinical case report

ABSTRACT Introduction: Purulent pericarditis is an inflammatory process in the pericardium caused by bacterial infection. If experienced during childhood and with untimely diagnosis, it has a high mortality rate. Case presentation: A 10-month-old infant was admitted to a high complexity pediatric hospital in the city of Bogotá D.C, Colombia, due to clinical symptoms including cough, respiratory distress and fever. A chest x-ray was taken showing cardiomegaly and multilobar pulmonary involvement. The echocardiogram showed global pericardial effusion managed with pericardiotomy, in which 50 mL of turbid fluid with whitish membranes was obtained. Cytochemical test revealed 2 600 mm3 leukocytes with 90% PMN and protein elevation. Purulent pericarditis was diagnosed based on imaging and laboratory findings. Treatment was initiated with ceftriaxone and clindamycin for four weeks, obtaining effective clinical and echocardiographic resolution. Discussion: The clinical presentation and imaging, paraclinical and electrocardiographic findings suggested purulent pericarditis as the first possibility. This diagnosis was confirmed considering the characteristics of the pericardial fluid, which was compatible with an exudate. Clinical resolution supported by antibiotic management corroborated the diagnosis, even though microbiological isolation was not obtained in cultures. Conclusion: Purulent pericarditis is a rare disease in pediatrics and has a high mortality rate. Making a timely diagnosis and administering early treatment are related to a better prognosis of this pathology.

RESUMEN Introducción. La pericarditis purulenta es un proceso inflamatorio del pericardio producto de una infección bacteriana. De no lograrse un diagnóstico oportuno, se convierte en una patología con alta mortalidad en la infancia. Presentación del caso. Lactante de 10 meses de edad que ingresó a un hospital pediátrico de alta complejidad en Bogotá D.C., Colombia, por un cuadro clínico dado por tos, dificultad respiratoria y fiebre. Se tomó una radiografía de tórax donde se observó cardiomegalia y compromiso neumónico multilobar. El ecocardiograma mostró un derrame pericárdico global que requirió pericardiotomía, en la cual se obtuvo 50 mL de líquido turbio con membranas blanquecinas. En la prueba citoquímica se encontraron 2 600mm3 leucocitos, polimorfonucleares del 90% y elevación de proteínas. Con los hallazgos de imagenología y laboratorio se hizo el diagnóstico de pericarditis purulenta, por lo que se inició tratamiento con ceftriaxona y clindamicina por 4 semanas, obteniendo una resolución clínica y ecocardiográfica efectiva. Discusión. La presentación clínica y los hallazgos imagenológicos, paraclínicos y electrocardiográficos sugirieron como primera posibilidad pericarditis purulenta, lo cual se confirmó por las características de líquido pericárdico, que era compatible con un exudado. La resolución clínica, apoyada por el manejo antibiótico y a pesar de no obtener aislamiento microbiológico en los cultivos, corroboró el diagnóstico. Conclusiones. La pericarditis purulenta es una enfermedad poco frecuente en pediatría pero con alta mortalidad. Realizar un diagnóstico oportuno sumado a un tratamiento tempano se relaciona con un mejor pronóstico de esta patología.

Paracrine cellular senescence exacerbates biliary injury and impairs regeneration.

Cellular senescence is a mechanism that provides an irreversible barrier to cell cycle progression to prevent undesired proliferation. However, under pathological circumstances, senescence can adversely affect organ function, viability and regeneration. We have developed a mouse model of biliary senescence, based on the conditional deletion of Mdm2 in bile ducts under the control of the Krt19 promoter, that exhibits features of biliary disease. Here we report that senescent cholangiocytes induce profound alterations in the cellular and signalling microenvironment, with recruitment of myofibroblasts and macrophages causing collagen deposition, TGFß production and induction of senescence in surrounding cholangiocytes and hepatocytes. Finally, we study how inhibition of TGFß-signalling disrupts the transmission of senescence and restores liver function. We identify cellular senescence as a detrimental mechanism in the development of biliary injury. Our results identify TGFß as a potential therapeutic target to limit senescence-dependent aggravation in human cholangiopathies.

An adaptive signaling network in melanoma inflammatory niches confers tolerance to MAPK signaling inhibition.

Mitogen-activated protein kinase (MAPK) pathway antagonists induce profound clinical responses in advanced cutaneous melanoma, but complete remissions are frustrated by the development of acquired resistance. Before resistance emerges, adaptive responses establish a mutation-independent drug tolerance. Antagonizing these adaptive responses could improve drug effects, thereby thwarting the emergence of acquired resistance. In this study, we reveal that inflammatory niches consisting of tumor-associated macrophages and fibroblasts contribute to treatment tolerance through a cytokine-signaling network that involves macrophage-derived IL-1ß and fibroblast-derived CXCR2 ligands. Fibroblasts require IL-1ß to produce CXCR2 ligands, and loss of host IL-1R signaling in vivo reduces melanoma growth. In tumors from patients on treatment, signaling from inflammatory niches is amplified in the presence of MAPK inhibitors. Signaling from inflammatory niches counteracts combined BRAF/MEK (MAPK/extracellular signal-regulated kinase kinase) inhibitor treatment, and consequently, inhibiting IL-1R or CXCR2 signaling in vivo enhanced the efficacy of MAPK inhibitors. We conclude that melanoma inflammatory niches adapt to and confer drug tolerance toward BRAF and MEK inhibitors early during treatment.

Detecting the Senescence-Associated Secretory Phenotype (SASP) by High Content Microscopy Analysis.

The diverse arrays of proteins secreted by senescent cells have been described to influence aging and to have both pro-tumorigenic and anti-tumorigenic influences on the surrounding microenvironment. Further characterization of these proteins, known as the senescence-associated secretory phenotype (SASP), and their regulators is required to understand and further manipulate such activities. The use of high-throughput technology allows us to obtain visual and quantitative data on a large number of samples quickly and easily. Not only is this an invaluable tool for conducting large-scale RNAi or compound screenings, but also allows rapid validation of candidates of interest. Here, we describe how we use the Widefield High-Content Analysis Systems to characterize the phenotypes of cells following modulation by potential regulators of Oncogene-Induced Senescence (OIS) by measuring numerous markers of senescence, including the SASP. This approach can be also used to screen for siRNA able to perturb the expression of SASP components during OIS.