Estudio de genes y proteínas de autofagia en tumores del sistema nervioso central

[ES] El glioblastoma multiforme (GBM) es el tumor maligno del sistema nervioso central más común en adultos. Se trata de un astrocitoma de grado IV con elevada heterogeneidad morfológica y genética caracterizado por tener una proliferación celular incontrolada, elevada capacidad de infiltración, angiogénesis robusta, resistencia a la apoptosis, propensión al desarrollo de focos necróticos y alta inestabilidad genómica. A pesar del creciente conocimiento de este tumor, tanto a nivel genético como molecular, el glioblastoma multiforme es uno de los tumores humanos más letales y el pronóstico de los pacientes continúa siendo desfavorable, con una supervivencia aproximada de un año tras el diagnóstico. La autofagia es un sistema de degradación intracelular altamente regulado por el cual proteínas de larga vida, orgánulos o incluso patógenos son absorbidos por vesículas de doble membrana llamadas autofagosomas y trasportados hasta el lisosoma, donde serán degradados. Tras esto, las macromoléculas resultantes serán translocadas al citosol donde se reciclarán. Se sabe que la autofagia desempeña un papel importante en el desarrollo de enfermedades, incluyendo enfermedades neurodegenerativas, cardiovasculares, inflamatorias, hepáticas, diabetes y cáncer, donde puede tanto promover como inhibir la tumorigénesis. Es por ello que la modulación del proceso autofágico, tanto para inhibirlo como para hiperactivarlo, podría ser una interesante opción terapéutica en tumores y, en particular, en glioblastoma multiforme. El principal objetivo de este trabajo ha sido determinar si la autofagia podría estar relacionada específicamente con el desarrollo y la evolución del glioblastoma multiforme y podría ser considerada una nueva diana terapéutica para esta enfermedad. Para ello hemos analizado diferentes variantes polimórficas en genes de autofagia como posibles factores relacionados con la susceptibilidad a padecer glioblastoma multiforme; hemos analizado la expresión de proteínas de la maquinaria autofágica en pacientes con glioblastomas y astrocitomas de bajo grado para profundizar en el conocimiento del proceso de autofagia y determinar posibles biomarcadores pronósticos y diagnósticos de dichos tumores que ayuden a predecir su desarrollo y evolución y, por último, hemos analizado el efecto de los fármacos moduladores de la autofagia Panobinostat®, cloroquina, metformina, paclitaxel, y las sinergias Panobinostat® + cloroquina, 5-aza-2’-desoxicitidina + cloroquina y 5-aza-2’-desoxicitidina + metformina en líneas tumorales de glioblastoma multiforme para determinar nuevas estrategias terapéuticas en el tratamiento de este tumor

Whole blood DNA methylation analysis reveals respiratory environmental traits involved in COVID-19 severity following SARS-CoV-2 infection

SARS-CoV-2 infection can cause an inflammatory syndrome (COVID-19) leading, in many cases, to bilateral pneumonia, severe dyspnea, and in ~5% of these, death. DNAmethylation is known to play an important role in the regulation of the immune processes behind COVID-19 progression, however it has not been studied in depth. In this study, we aim to evaluate the implication of DNA methylation in COVID-19 progression by means of a genome-wide DNA methylation analysis combined with DNA genotyping. The results reveal the existence of epigenomic regulation of functional pathways associated with COVID-19 progression andmediated by genetic loci.We find an environmental trait-related signature that discriminatesmild from severe cases and regulates, among other cytokines, IL-6 expression via the transcription factor CEBP. The analyses suggest that an interaction between environmental contribution, genetics, and epigenetics might be playing a role in triggering the cytokine storm described in the most severe cases.Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades of the regional government of AndaluciaEuropean Union through European Regional Development Fund CV20-10150Consejo Superior de Investigaciones cientificas CSIC-COV19-016/202020E155Junta de Castilla y Leon COVID 07.04.467B04.74011.0Consejeria de Salud y Familias of the regional government of Andalucia PECOVID-0072-2020Instituto de Salud Carlos III (ISCIII, Spanish Health Ministry) through the Sara Borrell subprogram CD18/00153Programa Estrategico Instituto de Biologia y Genetica Molecular, IBGM excellence programme CLU-2029-02 CCVC848

Actividad antimicobacteriana de terpenos

RESUMEN INTRODUCCIÓN: La tuberculosis (TB), causada por Mycobacterium tuberculosis es la mayor causa de mortalidad mundial por un único agente patógeno. Asimismo, un gran número de micobacterias no tuberculosas, especialmente M. avium, M. intracellulare y M. chelonae, causan infecciones oportunistas en pacientes con SIDA. Muchos terpenos poseen actividad biológica y se emplean en el tratamiento de diversas enfermedades, razón que los hace fuente de moléculas promisorias. OBJETIVO: El objetivo del presente estudio fue determinar la actividad antimicobacteriana de 16 terpenos contra M. tuberculosis H37Rv y un aislamiento clínico de M. chelonae.  MATERIALES Y MÉTODOS: Se obtuvo la concentración mínima inhibitoria (CMI) de los mismos y se realizaron curvas de letalidad para establecer actividad bactericida, empleando una técnica de macrodilución en caldo estandarizada para este tipo de compuestos volátiles. RESULTADOS: Los terpenos con menor valor de CMI fueron timol y carvacrol, con concentraciones de 125-250 µg/mL, y actividad bactericida contra los dos microorganismos. Geraniol, mirceno, p-cimeno, alfa-pineno, presentaron valores de CMI entre 250 y 500 µg/mL. CONCLUSIONES: Algunos terpenos han presentado actividad importante contra microorganismos del género Mycobacterium, sin embargo los valores de CMI obtenidos no explican el efecto antimicrobiano presentado por el aceite completo, se requiere evaluar las interacciones de sinergismo y/o antagonismo entre los terpenos para determinar los componentes responsables de la acción farmacológica. Palabras clave: Actividad antimicobacteriana, terpenos, tuberculosis ABSTRACT INTRODUCTION: Tuberculosis (TB) caused by Mycobacterium tuberculosis is the major source of global mortality from a single pathogen. Moreover, a large number of nontuberculous mycobacteria, especially M. avium, M. intracellulare and M. chelonae, cause opportunistic infection in AIDS patients. Terpenes, possess a wide spectrum of biological activity and are used in the treatment of various diseases, reason that makes them a source of promising molecules. OBJECTIVE: To determine the antimycobacterial activity of 16 terpenes against M. tuberculosis H37Rv and a clinical isolate of M. chelonae. MATERIALS AND METHODS: Minimum Inhibitory Concentration (MIC) values of terpenes were obtained by macrodilution colorimetric method standardized for this kind of volatile compounds, time kill curves were determined to establish bactericidal activity using CLSIM26-A method. RESULTS: Terpenes with the lower MIC values were thymol and carvacrol at concentrations of 125-250 µg/mL, showing bactericidal activity against both microorganisms. Geraniol, myrcene, p-cymene, and alpha - pinene, showed MIC values ranging between 250-500 µg/mL. CONCLUSIÓN: Some terpenes have presented signifcant activity against microorganisms of the Mycobacterium genus, but the MIC values obtained do not explain the antimicrobial effect showed by the complete oil, is necessary to assess the interactions of synergism and / or antagonism between the terpenes to determine the components responsible for Pharmacological.  Keywords: Antimycobacterial activity, terpenes, tuberculosi

Actividad antimicobacteriana de terpenos

RESUMEN INTRODUCCIÓN: La tuberculosis (TB), causada por Mycobacterium tuberculosis es la mayor causa de mortalidad mundial por un único agente patógeno. Asimismo, un gran número de micobacterias no tuberculosas, especialmente M. avium, M. intracellulare y M. chelonae, causan infecciones oportunistas en pacientes con SIDA. Muchos terpenos poseen actividad biológica y se emplean en el tratamiento de diversas enfermedades, razón que los hace fuente de moléculas promisorias. OBJETIVO: El objetivo del presente estudio fue determinar la actividad antimicobacteriana de 16 terpenos contra M. tuberculosis H37Rv y un aislamiento clínico de M. chelonae.  MATERIALES Y MÉTODOS: Se obtuvo la concentración mínima inhibitoria (CMI) de los mismos y se realizaron curvas de letalidad para establecer actividad bactericida, empleando una técnica de macrodilución en caldo estandarizada para este tipo de compuestos volátiles. RESULTADOS: Los terpenos con menor valor de CMI fueron timol y carvacrol, con concentraciones de 125-250 µg/mL, y actividad bactericida contra los dos microorganismos. Geraniol, mirceno, p-cimeno, alfa-pineno, presentaron valores de CMI entre 250 y 500 µg/mL. CONCLUSIONES: Algunos terpenos han presentado actividad importante contra microorganismos del género Mycobacterium, sin embargo los valores de CMI obtenidos no explican el efecto antimicrobiano presentado por el aceite completo, se requiere evaluar las interacciones de sinergismo y/o antagonismo entre los terpenos para determinar los componentes responsables de la acción farmacológica. Palabras clave: Actividad antimicobacteriana, terpenos, tuberculosis ABSTRACT INTRODUCTION: Tuberculosis (TB) caused by Mycobacterium tuberculosis is the major source of global mortality from a single pathogen. Moreover, a large number of nontuberculous mycobacteria, especially M. avium, M. intracellulare and M. chelonae, cause opportunistic infection in AIDS patients. Terpenes, possess a wide spectrum of biological activity and are used in the treatment of various diseases, reason that makes them a source of promising molecules. OBJECTIVE: To determine the antimycobacterial activity of 16 terpenes against M. tuberculosis H37Rv and a clinical isolate of M. chelonae. MATERIALS AND METHODS: Minimum Inhibitory Concentration (MIC) values of terpenes were obtained by macrodilution colorimetric method standardized for this kind of volatile compounds, time kill curves were determined to establish bactericidal activity using CLSIM26-A method. RESULTS: Terpenes with the lower MIC values were thymol and carvacrol at concentrations of 125-250 µg/mL, showing bactericidal activity against both microorganisms. Geraniol, myrcene, p-cymene, and alpha - pinene, showed MIC values ranging between 250-500 µg/mL. CONCLUSIÓN: Some terpenes have presented signifcant activity against microorganisms of the Mycobacterium genus, but the MIC values obtained do not explain the antimicrobial effect showed by the complete oil, is necessary to assess the interactions of synergism and / or antagonism between the terpenes to determine the components responsible for Pharmacological.  Keywords: Antimycobacterial activity, terpenes, tuberculosi

Scoring personalized molecular portraits identify Systemic Lupus Erythematosus subtypes and predict individualized drug responses, symptomatology and disease progression

Objectives Systemic Lupus Erythematosus is a complex autoimmune disease that leads to significant worsening of quality of life and mortality. Flares appear unpredictably during the disease course and therapies used are often only partially effective. These challenges are mainly due to the molecular heterogeneity of the disease, and in this context, personalized medicine-based approaches offer major promise. With this work we intended to advance in that direction by developing MyPROSLE, an omic-based analytical workflow for measuring the molecular portrait of individual patients to support clinicians in their therapeutic decisions. Methods Immunological gene-modules were used to represent the transcriptome of the patients. A dysregulation score for each gene-module was calculated at the patient level based on averaged z-scores. Almost 6100 Lupus and 750 healthy samples were used to analyze the association among dysregulation scores, clinical manifestations, prognosis, flare and remission events and response to Tabalumab. Machine learning-based classification models were built to predict around 100 different clinical parameters based on personalized dysregulation scores. Results MyPROSLE allows to molecularly summarize patients in 206 gene-modules, clustered into nine main lupus signatures. The combination of these modules revealed highly differentiated pathological mechanisms. We found that the dysregulation of certain gene-modules is strongly associated with specific clinical manifestations, the occurrence of relapses or the presence of long-term remission and drug response. Therefore, MyPROSLE may be used to accurately predict these clinical outcomes. Conclusions MyPROSLE (https://myprosle.genyo.es) allows molecular characterization of individual Lupus patients and it extracts key molecular information to support more precise therapeutic decisions.PID2020-119032RB-I00 supported by MCIN/AEI/10.13039/501100011033FEDER and the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No 831434 (3TR)European Union’s Horizon 2020EFPIAFEDER/Junta de Andalucía-Consejer’a de Transformación Económica, Industria, Conocimiento y Universidades (grants P20_00335 and B-CTS-40-UGR20)‘Consejería de Transformación Económica, Industria, Conocimiento y Universidades’ (CTEICU)European Union through the European Social Fund (ESF) named ‘Andalucía se mueve con Europa”Andalusian ESF Operational Program 2014–2020ISCIII CD18/00149Ministerio de Universidades (Spain’s Government) and the European Union – NextGenerationE

Hyperphosphatemia-Induced Oxidant/Antioxidant Imbalance Impairs Vascular Relaxation and Induces Inflammation and Fibrosis in Old Mice

Aging impairs vascular function, but the mechanisms involved are unknown. The aim of this study was to analyze whether aging-related hyperphosphatemia is implied in this effect by elucidating the role of oxidative stress. C57BL6 mice that were aged 5 months (young) and 24 months (old), receiving a standard (0.6%) or low-phosphate (0.2%) diet, were used. Isolated mesenteric arteries from old mice showed diminished endothelium-dependent vascular relaxation by the down-regulation of NOS3 expression, increased inflammation and increased fibrosis in isolated aortas, compared to those isolated from young mice. In parallel, increased Nox4 expression and reduced Nrf2, Sod2-Mn and Gpx1 were found in the aortas from old mice, resulting in oxidant/antioxidant imbalance. The low-phosphate diet improved vascular function and oxidant/antioxidant balance in old mice. Mechanisms were analyzed in endothelial (EC) and vascular smooth muscle cells (SMCs) treated with the phosphate donor ss-glycerophosphate (BGP). In EC, BGP increased Nox4 expression and ROS production, which reduced NOS3 expression via NF kappa B. BGP also increased inflammation in EC. In SMC, BGP increased Collagen I and fibronectin expression by priming ROS production and NF kappa B activity. In conclusion, hyperphosphatemia reduced endothelium-dependent vascular relaxation and increased inflammation and vascular fibrosis through an impairment of oxidant/antioxidant balance in old mice. A low-phosphate diet achieved improvements in the vascular function in old mice

Actividad in vitro anti-candida y anti-aspergillus de aceites esenciales de plantas de la familia piperaceae

Plants of the Piperaceae family have been used with medicinal purposes in different regions around the world. In vitro antifungal activity of 10 essential oils from species of Piper genus were measured by determination of minimum inhibitory concentration (MIC) against A. flavus fungi ATCC 204304 and A. fumigatus ATCC 204305 following the standard protocol CLSI M38-A. In addition, Candida krusei ATCC 6258 and C. parapsilosis ATCC 22019 were evaluated with modified protocol M27-A2. The essential oil with the most antifungal activity was Piper sanctifelisis for C. krusei (CMI: 125 ìg/mL).Plantas de la familia Piperaceae se han utilizado con fines curativos en diferentes regiones del mundo. El objetivo del presente fue evaluar la actividad antifungica in vitro de 10 aceites esenciales provenientes de especies del genero Piper, mediante la determinacion de la concentracion minima inhibitoria (CMI), con las cepas Aspergillus flavus ATCC 204304 y A. fumigatus ATCC 20430, siguiendo el protocolo CLSI M38-A. Ademas, se evaluaron las cepas Candida krusei ATCC 6258 y Candida parapsilosis ATCC 22019 segun el protocolo M27-A2 modificado. El aceite esencial con mayor actividad fue el de P. sanctifelisis para C. krusei (CMI:125 �Êg/mL)

Integrative epigenomics in Sjögren¿s syndrome reveals novel pathways and a strong interaction between the HLA, autoantibodies and the interferon signature.

Primary Sjögren’s syndrome (SS) is a systemic autoimmune disease characterized by lymphocytic infiltration and damage of exocrine salivary and lacrimal glands. The etiology of SS is complex with environmental triggers and genetic factors involved. By conducting an integrated multi-omics study, we confirmed a vast coordinated hypomethylation and overexpression effects in IFN-related genes, what is known as the IFN signature. Stratified and conditional analyses suggest a strong interaction between SS-associated HLA genetic variation and the presence of Anti-Ro/SSA autoantibodies in driving the IFN epigenetic signature and determining SS. We report a novel epigenetic signature characterized by increased DNA methylation levels in a large number of genes enriched in pathways such as collagen metabolism and extracellular matrix organization. We identified potential new genetic variants associated with SS that might mediate their risk by altering DNA methylation or gene expression patterns, as well as disease-interacting genetic variants that exhibit regulatory function only in the SS population. Our study sheds new light on the interaction between genetics, autoantibody profiles, DNA methylation and gene expression in SS, and contributes to elucidate the genetic architecture of gene regulation in an autoimmune population.Funding for the preparation of this manuscript has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement nº 115,565, resources composed of the financial contribution from the European Union's Seventh Framework Program (FP7/2007-2013) and the EFPIA companies’ in kind contribution. MT is supported by a Spanish grant from Health Department, Junta de Andalucía (PI/0017/2016) and through the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement No 806975. This Joint Undertaking receives support from the European Union’s Horizon 2020 research and innovation programme and EFPIA. EC-M was funded by the Postdoctoral Training Subprogramme Juan de la Cierva-Ministry of Economy and Competitiveness (FJCI_2014_20652). We thank Ralf Lesche for the production of RNASeq data and Marc Torres Ciuró for design support.Peer reviewe