Interaction modulation through arrays of clustered methyl-arginine protein modifications

Systematic analysis of human arginine methylation identifies two distinct signaling modes;either isolated modifications akin to canonical post-translational modification regulation, or clustered arrays within disordered protein sequence. Hundreds of proteins contain these methyl-arginine arrays and are more prone to accumulate mutations and more tightly expression-regulated than dispersed methylation targets. Arginines within an array in the highly methylated RNA-binding protein synaptotagmin binding cytoplasmic RNA interacting protein (SYNCRIP) were experimentally shown to function in concert, providing a tunable protein interaction interface. Quantitative immunoprecipitation assays defined two distinct cumulative binding mechanisms operating across 18 proximal arginine-glycine (RG) motifs in SYNCRIP. Functional binding to the methyltransferase PRMT1 was promoted by continual arginine stretches, whereas interaction with the methyl-binding protein SMN1 was arginine content-dependent irrespective of linear position within the unstructured region. This study highlights how highly repetitive modifiable amino acid arrays in low structural complexity regions can provide regulatory platforms, with SYNCRIP as an extreme example how arginine methylation leverages these disordered sequences to mediate cellular interactions

Differential Regulation of Gene Expression in Lung Cancer Cells by Diacyglycerol-Lactones and a Phorbol Ester Via Selective Activation of Protein Kinase C Isozymes

Despite our extensive knowledge on the biology of protein kinase C (PKC) and its involvement in disease, limited success has been attained in the generation of PKC isozyme-specifc modulators acting via the C1 domain, the binding site for the lipid second messenger diacylglycerol (DAG) and the phorbol ester tumor promoters. Synthetic eforts had recently led to the identifcation of AJH-836, a DAG-lactone with preferential afnity for novel isozymes (nPKCs) relative to classical PKCs (cPKCs). Here, we compared the ability of AJH-836 and a prototypical phorbol ester (phorbol 12-myristate 13-acetate, PMA) to induce changes in gene expression in a lung cancer model. Gene profling analysis using RNA-Seq revealed that PMA caused major changes in gene expression, whereas AJH-836 only induced a small subset of genes, thus providing a strong indication for a major involvement of cPKCs in their control of gene expression. MMP1, MMP9, and MMP10 were among the genes most prominently induced by PMA, an efect impaired by RNAi silencing of PKCα, but not PKCδ or PKCε. Comprehensive gene signature analysis and bioinformatics eforts, including functional enrichment and transcription factor binding site analyses of dysregulated genes, identifed major diferences in pathway activation and transcriptional networks between PMA and DAG-lactones. In addition to providing solid evidence for the diferential involvement of individual PKC isozymes in the control of gene expression, our studies emphasize the importance of generating targeted C1 domain ligands capable of diferentially regulating PKC isozyme-specifc function in cellular models.Centro de Investigaciones Inmunológicas Básicas y Aplicada

CIBERER : Spanish national network for research on rare diseases: A highly productive collaborative initiative

Altres ajuts: Instituto de Salud Carlos III (ISCIII); Ministerio de Ciencia e Innovación.CIBER (Center for Biomedical Network Research; Centro de Investigación Biomédica En Red) is a public national consortium created in 2006 under the umbrella of the Spanish National Institute of Health Carlos III (ISCIII). This innovative research structure comprises 11 different specific areas dedicated to the main public health priorities in the National Health System. CIBERER, the thematic area of CIBER focused on rare diseases (RDs) currently consists of 75 research groups belonging to universities, research centers, and hospitals of the entire country. CIBERER's mission is to be a center prioritizing and favoring collaboration and cooperation between biomedical and clinical research groups, with special emphasis on the aspects of genetic, molecular, biochemical, and cellular research of RDs. This research is the basis for providing new tools for the diagnosis and therapy of low-prevalence diseases, in line with the International Rare Diseases Research Consortium (IRDiRC) objectives, thus favoring translational research between the scientific environment of the laboratory and the clinical setting of health centers. In this article, we intend to review CIBERER's 15-year journey and summarize the main results obtained in terms of internationalization, scientific production, contributions toward the discovery of new therapies and novel genes associated to diseases, cooperation with patients' associations and many other topics related to RD research

Análisis del papel de beta2-quimerina en cáncer de mama

Beta2-quimerina es una proteína con actividad GAP específica de Rac. Rac es una RhoGTPasa que regula múltiples procesos celulares,muchos de los cuales se encuentran alterados en cáncer de mama, como son proliferación, supervivencia, motilidad y desdiferenciación celular. Evidencias previas a este estudio han demostrado la reducción de beta2-quimerína en cáncer de mama,sugiriendo un papel para esta proteína de supresor tumoral en ésta patología. La inactivación de la proteína Rac tendría como resultado un incremento en la actividad de Rac. Para evaluar el papel de beta2-quimerina en el desarrollo del cáncer de mama in vivo se generó un modelo de ratón con defecto de expresión de esta isoforma en ratones transgénicos MMTV/Neu en el que se sobreexpresa el receptor ErbB2

Efecto antileucémico de derivados cannabinoides: un nuevo grupo terapéutico en LMA

2 p.-3 fig.Los cannabinoides, componentes activos del cannabis, se emplean como tratamiento paliativo en pacientes con cáncer en virtud de su efecto antiemético y analgésico. Además, hay evidencias de que estos compuestos inhiben el crecimiento de algunas células tumorales en animales de laboratorio. Estos compuestos actúan uniéndose a receptores específicos. Los principales receptores son el CB1 (mayoritario en SNC) y CB2 (en tejido hematopoyético). El objetivo de este estudio fue confirmar el efecto terapéutico de derivados cannabinoides sintéticos específicos del receptor CB2 en LMA.Peer reviewe

Differential Regulation of Gene Expression in Lung Cancer Cells by Diacyglycerol-Lactones and a Phorbol Ester Via Selective Activation of Protein Kinase C Isozymes

Abstract Despite our extensive knowledge on the biology of protein kinase C (PKC) and its involvement in disease, limited success has been attained in the generation of PKC isozyme-specific modulators acting via the C1 domain, the binding site for the lipid second messenger diacylglycerol (DAG) and the phorbol ester tumor promoters. Synthetic efforts had recently led to the identification of AJH-836, a DAG-lactone with preferential affinity for novel isozymes (nPKCs) relative to classical PKCs (cPKCs). Here, we compared the ability of AJH-836 and a prototypical phorbol ester (phorbol 12-myristate 13-acetate, PMA) to induce changes in gene expression in a lung cancer model. Gene profiling analysis using RNA-Seq revealed that PMA caused major changes in gene expression, whereas AJH-836 only induced a small subset of genes, thus providing a strong indication for a major involvement of cPKCs in their control of gene expression. MMP1, MMP9, and MMP10 were among the genes most prominently induced by PMA, an effect impaired by RNAi silencing of PKCα, but not PKCδ or PKCε. Comprehensive gene signature analysis and bioinformatics efforts, including functional enrichment and transcription factor binding site analyses of dysregulated genes, identified major differences in pathway activation and transcriptional networks between PMA and DAG-lactones. In addition to providing solid evidence for the differential involvement of individual PKC isozymes in the control of gene expression, our studies emphasize the importance of generating targeted C1 domain ligands capable of differentially regulating PKC isozyme-specific function in cellular models

Single residue in CD28-costimulated CAR-T cells limits long-term persistence and antitumor durability

Chimeric antigen receptor–T (CAR-T) cell therapies can eliminate relapsed and refractory tumors, but the durability of antitumor activity requires in vivo persistence. Differential signaling through the CAR costimulatory domain can alter the T cell metabolism, memory differentiation, and influence long-term persistence. CAR-T cells costimulated with 4-1BB or ICOS persist in xenograft models but those constructed with CD28 exhibit rapid clearance. Here, we show that a single amino acid residue in CD28 drove T cell exhaustion and hindered the persistence of CD28-based CAR-T cells and changing this asparagine to phenylalanine (CD28-YMFM) promoted durable antitumor control. In addition, CD28-YMFM CAR-T cells exhibited reduced T cell differentiation and exhaustion as well as increased skewing toward Th17 cells. Reciprocal modification of ICOS-containing CAR-T cells abolished in vivo persistence and antitumor activity. This finding suggests modifications to the costimulatory domains of CAR-T cells can enable longer persistence and thereby improve antitumor response

Cannabinoids derivatives modify the pattern of sphingolipids in acute myeloid leukemia cells and produce a potent anti-leukemic effect

2 p. 22nd Congress of the European Hematology Association. Madrid, Spain, June 22 - 25, 2017Endocannabinoid system is a set of ligands, receptors and endogenous enzymes which modulate a variety of physiological effects. There are two well-characterized cannabinoid receptors, CB1 (mainly expressed in Central Nervous System) and CB2 (mainly in hematopoietic cells). Here, we tested the effect of the cannabinoid WIN-55 212-2 in acute myeloid leukemia (AML) in vitro and in vivo and studied the molecular signaling pathways involved in this effect, specially the role of sphingolipids. Moreover, we synthesized a new family of twelve cannabinoids that are specific to CB2 receptor.Peer reviewe

Indoor air quality in a training centre used for sports practice

Background One of the measures for controlling the coronavirus disease 2019 (COVID-19) pandemic was the mass closure of gyms. This measure leads us to determine the differences between indoor and outdoor air quality. That is why the objective of this study was to analyse the indoor air quality of a sports centre catering to small groups and rehabilitation. Methods The study was conducted in a single training centre, where 26 measurements were taken in two spaces (indoors and outdoors). The air quality index, temperature, relative humidity, total volatile compounds, carbon monoxide, ozone, formaldehyde, carbon dioxide, and particulate matter were measured indoors and outdoors using the same protocol and equipment. These measurements were taken twice, once in the morning and once in the afternoon, with all measurements made at the same time, 10 am and 6 pm, respectively. Additionally, four determinations of each variable were collected during each shift, and the number of people who had trained in the room and the number of trainers were counted. Results In the different variables analysed, the results show that CO2 and RH levels are higher indoors than outdoors in both measurement shifts. Temperatures are higher outside than inside and, in the evening, than in the morning. TVOC, AQI and PM show less variation, although they are higher outdoors in the morning. CO is highest indoors. HCHO levels are almost negligible and do not vary significantly, except for a slight increase in the afternoon outside. Ozone levels are not significant. All the variables showed practically perfect reliability in all the measurements, except for ozone measured outside in the morning. On the other hand, the variables exhibit variations between indoors and outdoors during the morning and afternoon, except for the three types of PM. Also, the data show that all the main variables measured inside the sports training centre are similar between morning and afternoon. However, outside, temperature, relative humidity and HCHO levels show significant differences between morning and afternoon while no differences are observed for the other variables. Conclusion The indoor air quality of the training centre assessed was good and met current regulations; some of its components even exhibited better levels than fresh air. This article is the first to measure indoor air quality in a sports training centre catering to rehabilitation and small groups

Development of Novel Murine BRAF^V600E-Driven Papillary Thyroid Cancer Cell Lines for Modeling of Disease Progression and Preclinical Evaluation of Therapeutics

The Cancer Genome Atlas study in thyroid cancer exposed the genomic landscape of ~500 PTCs and revealed BRAFV600E-mutant tumors as having different prognosis, contrasting indolent cases and those with more invasive disease. Here, we describe the generation and characterization of six novel BRAFV600E-driven papillary thyroid cancer (PTC) cell lines established from a BrafV600E+/−/Pten+/−/TPO-Cre mouse model that spontaneously develop thyroid tumors. The novel cell lines were obtained from animals representing a range of developmental stages and both sexes, with the goal of establishing a heterogeneous panel of PTC cell lines sharing a common driver mutation. These cell lines recapitulate the genetics and diverse histopathological features of BRAFV600E-driven PTC, exhibiting differing degrees of growth, differentiation, and invasive potential that may help define mechanisms of pathogenesis underlying the heterogeneity present in the patient population. We demonstrate that these cell lines can be used for a variety of in vitro applications and can maintain the potential for in vivo transplantation into immunocompetent hosts. We believe that these novel cell lines will provide powerful tools for investigating the molecular basis of thyroid cancer progression and will lead to the development of more personalized diagnostic and treatment strategies for BRAFV600E-driven PTC