Terapia con miRNAs en la enfermedad cardiovascular y envejecimiento: descubrimiento de nuevos candidatos

Cardiovascular diseases are, as of now, the main cause of death in the developed world. Aging is amain risk factor determining their development. In most of the cases, pharmacological treatments donot directly asses the problem and have only palliative effects, which decreases quality life of thepatient and increase dependence on the treatment to follow a normal lifestyle. Therefore, looking fornovel effective therapies to treat the cause of the disease will be crucial in the near future.microRNAs (miRNA) have been proved to be master regulators of many biological processes that takeplaces in the cell, since they are capable of regulating gene expression. Studies in vitro and in vivo haveassociated dysregulation of the miRNAs expression with pathology in cardiovascular disease and aging,thus postulating them as possible therapeutic targets.As result, miRNAs-mimics and anti-miRs have emerged as a new promising therapeutic approach tocorrect gene expression alterations regulated by miRNAs that have been seen in disease. Here, adetailed revision of the application of miRNA-based therapies to treat CVD is made. Although, currentlythere are only 2 studies that have reached clinical trials, many preclinical studies have publishedencouraging results.Continuing previous work of the group BSICoS, a second aim of this master’s thesis is to develop toolsfor the study of the interaction of age-related miRNAs (BIO-AGEmiRNA) with their target genes inrelation to cardiac function using human iPSC-derived cardiomyocytes. A inducible expression vectorwas created to be expressed mature miRNAs, specifically miR-4435, downstream of a fluorescentreporter gene. The vector was tested in vitro in HEK293 cells and proper expression of the reporterwas observed. However, expression of the mature miR-4435, and therefore proper processing of thepre-miRNA, could not be confirmed due to technical issues. Future work will continue to validate theremaining BIO-AGE-miRNAs in vitro, first in HEk393 and then in iPSC-derived cardiomyocytes were themolecular mechanism and functionality of the BIO-AGEmiRNAs will be investigated. The developedvector will be most certainly a useful tool to complete this future work.<br /

Análisis de los microRNA en enfermedades cardiovasculares y envejecimiento cardiaco

Cardiovascular diseases (CVD) are one of the main causes of death that affect worldwide. Thereare several risk factors for the development of CVD, among them, aging is important, whichimplies the anatomical and functional deterioration of the heart. CVD constitute a great burdenin advanced societies. There is a need for the development of therapies and non-invasiveprognostic and diagnostic biomarkers to effectively tackle them. microRNAs (miRNAs) are smallRNAs known to regulate main biological process. For this reason, miRNAs have been proposedas a powerful therapeutic targets or biomarkers.Here the role of miRNAs in the development of CVD and cardiac aging has been reviewed. Inaddition, the translation level of results in animal models to human has been investigated. Asresult, a complete compilation of miRNAs that contribute to CVD and cardiac aging is offered.This literature compilation points out that the great majority of the described miRNAs remainunstudied in human. Based in the high degree of miRNA sequence conservation between speciesand their target sequences, similar functions could be expected in human. Yet, otherphysiological differences in comparison with animal models could suggest the contrary. Toassess the contribution of miRNA in cardiac related pathophysiology, cardiomyocytes derivedfrom human induced pluripotent stem cells (iPSC-CMs) emerge as a model for their studyavoiding this translation difficulties.In addition, the BSICoS group has built a bioinformatic regulatory network controlled by miRNAsassociated with biological age (BIO-AGEmiRNAs) of the human myocardium. Experimentalvalidation of the interactions within this network is required. A new cost-effective validation toolhas been built using fluorescence. Last, luciferase reporter assays (DualGlo assay) are used tostudy interaction of cardiac-related genes with miR-3916. As a result, positive interaction trendshave been observed for CASQ2, ACTN2 and DSP with miR-3916. This fact highlights the potentialof miRNAs in the regulation of genes associated with human cardiac aging and CVD and theirpowerful value as therapeutic targets.<br /

Envejecimiento cardiaco humano: transcriptómica y desarrollo de modelos celulares de envejecimiento

Pese a la relevancia de la edad como factor de riesgo de las enfermedades cardiovasculares, sus bases moleculares y funcionales están poco caracterizadas en humanos. Para mejorar esta comprensión hemos estudiado su dinámica transcripcional cronológica y biológica en ventrículo izquierdo humano y hemos desarrollado un modelo de envejecimiento celular cardiaco

microRNAs en envejecimiento cardiaco humano: dianas terapéuticas y biomarcadores

Cardiovascular diseases (CVD) are the leading cause of death globally, being age one of the most important predisposing risk factor. MicroRNAs (miRNAs) are key post-transcriptional regulators of multiple biological processes, including cardiac aging. MiRNAs are nowadays emerging therapeutics. Also, miRNAs are cutting-edge biomarkers, as they can be actively or passively released from tissues to body fluids (i.e. blood) acting as indicators of physiological and pathophysiological processes in tissues. However, the role of miRNAs in age-related cardiac remodelling is poorly understood in humans. This limits their use as anti-aging interventions, to reduce the risk of CVD, but also as biomarkers of the true physiological state of the heart in relation to age (biological age indicators). Our group recently described the miRNome dynamics with age, chronological and biological, of the human left ventricle. We identified a number of miRNAs related to biological age (BIO-AGEmiRNAs) and bioinformatically established their downstream gene regulation network. Also, a number of cardiac-enriched BIO-AGEmiRNAs were described. Here, we aim to validate the BIO-AGEmiRNA regulatory network and to describe miRNA indicators of the biological age of the heart.Las enfermedades cardiovasculares (ECV) son la principal causa de muerte globalmentem siendo la edad uno de los factores de riesgo más importantes. Los microRNA(miRNA) son reguladores postranscripcionales clave de múltiples procesos biológicos, entre ellos el envejecimiento cardíaco. Los miRNA son hoy en día dianas terapéuticas emergente. Además, los miRNA son biomarcadores de vanguardia, ya que pueden ser liberados activa o pasiva de los tejidos a los fluidos corporales (es decir, la sangre) actuando como indicadores de procesos fisiológicos y fisiopatológicos en los tejidos. Sin embargo, el papel de los miRNA en la remodelación cardiaca relacionada con la edad esta escasamente estudiado en humanos. Esto limita su uso como intervenciones antienvejecimiento, para reducir el riesgo de ECV, pero también como biomarcadores del verdadero estado fisiológico del corazón en relación con la edad (indicadores de edad biológica). Nuestro grupo ha descrito recientemente la dinámica de expresión de los miRNAs con la edad, cronológica y biológica, del ventrículo izquierdo humano. Identificamos una serie de miRNAs relacionados con la edad biológica (BIO-AGEmiRNAs) y establecimos bioinformáticamente su red de regulación génica. Asimismo, se describió una serie de BIO-AGEmiRNAs enriquecidos en el corazón. Nuestro objetivo es validar la red reguladora BIO-AGEmiRNA y describir los miRNA indicadores de la edad biológica del corazón. &nbsp

Minimally invasive system to reliably characterize ventricular electrophysiology from living donors

Cardiac tissue slices preserve the heterogeneous structure and multicellularity of the myocardium and allow its functional characterization. However, access to human ventricular samples is scarce. We aim to demonstrate that slices from small transmural core biopsies collected from living donors during routine cardiac surgery preserve structural and functional properties of larger myocardial specimens, allowing accurate electrophysiological characterization. In pigs, we compared left ventricular transmural core biopsies with transmural tissue blocks from the same ventricular region. In humans, we analyzed transmural biopsies and papillary muscles from living donors. All tissues were vibratomesliced. By histological analysis of the transmural biopsies, we showed that tissue architecture and cellular organization were preserved. Enzymatic and vital staining methods verifed viability. Optically mapped transmembrane potentials confrmed that action potential duration and morphology were similar in pig biopsies and tissue blocks. Action potential morphology and duration in human biopsies and papillary muscles agreed with published ranges. In both pigs and humans, responses to increasing pacing frequencies and β-adrenergic stimulation were similar in transmural biopsies and larger tissues. We show that it is possible to successfully collect and characterize tissue slices from human myocardial biopsies routinely extracted from living donors, whose behavior mimics that of larger myocardial preparations both structurally and electrophysiologically.Fil: Oliván Viguera, Aida. Universidad de Zaragoza; EspañaFil: Pérez Zabalza, María. Universidad de Zaragoza; EspañaFil: García Mendívil, Laura. Universidad de Zaragoza; EspañaFil: Mountris, Konstantinos A.. Universidad de Zaragoza; EspañaFil: Orós Rodrigo, Sofía. Universidad de Zaragoza; EspañaFil: Ramos Marquès, Estel. Universidad de Zaragoza; EspañaFil: Vallejo Gil, José María. University Hospital Miguel Servet; EspañaFil: Fresneda Roldán, Pedro Carlos. University Hospital Miguel Servet; EspañaFil: Fañanás Mastral, Javier. University Hospital Miguel Servet; EspañaFil: Vázquez Sancho, Manuel. University Hospital Miguel Servet; EspañaFil: Matamala Adell, Marta. University Hospital Miguel Servet; EspañaFil: Sorribas Berjón, Fernando. University Hospital Miguel Servet; EspañaFil: Bellido Morales, Javier André. University Hospital Miguel Servet; EspañaFil: Mancebón Sierra, Francisco Javier. University Hospital Miguel Servet; EspañaFil: Vaca Núñez, Alexánder Sebastián. University Hospital Miguel Servet; EspañaFil: Ballester Cuenca, Carlos. University Hospital Miguel Servet; EspañaFil: Marigil, Miguel Ángel. Hospital San Jorge; EspañaFil: Pastor, Cristina. Aragón Institute of Health Sciences; EspañaFil: Ordovás, Laura. Aragón Agency for Research and Development; España. Universidad de Zaragoza; EspañaFil: Köhler, Ralf. Aragón Institute of Health Sciences; España. Aragón Agency for Research and Development; EspañaFil: Diez, Emiliano Raúl. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Cátedra de Fisiología Humana Normal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Pueyo, Esther. Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina; España. Universidad de Zaragoza; Españ

Chronological and biological aging of the human left ventricular myocardium: Analysis of microRNAs contribution

Aging is the main risk factor for cardiovascular diseases. In humans, cardiac aging remains poorly characterized. Most studies are based on chronological age (CA) and disregard biological age (BA), the actual physiological age (result of the aging rate on the organ structure and function), thus yielding potentially imperfect outcomes. Deciphering the molecular basis of ventricular aging, especially by BA, could lead to major progresses in cardiac research. We aim to describe the transcriptome dynamics of the aging left ventricle (LV) in humans according to both CA and BA and characterize the contribution of microRNAs, key transcriptional regulators. BA is measured using two CA-associated transcriptional markers: CDKN2A expression, a cell senescence marker, and apparent age (AppAge), a highly complex transcriptional index. Bioinformatics analysis of 132 LV samples shows that CDKN2A expression and AppAge represent transcriptomic changes better than CA. Both BA markers are biologically validated in relation to an aging phenotype associated with heart dysfunction, the amount of cardiac fibrosis. BA-based analyses uncover depleted cardiac-specific processes, among other relevant functions, that are undetected by CA. Twenty BA-related microRNAs are identified, and two of them highly heart-enriched that are present in plasma. We describe a microRNA-gene regulatory network related to cardiac processes that are partially validated in vitro and in LV samples from living donors. We prove the higher sensitivity of BA over CA to explain transcriptomic changes in the aging myocardium and report novel molecular insights into human LV biological aging. Our results can find application in future therapeutic and biomarker research

Construyendo circuitos neuronales

Uno de los criterios de evaluación contemplados en el currículum del tercer curso de la ESO en la asignatura de Biología y Geología es explicar la misión integradora del sistema nervioso ante diferentes estímulos y describir su funcionamiento. La actividad presentada tiene como objetivo el aprendizaje del funcionamiento de parte del sistema nervioso a nivel celular. Se usan modelos de órganos y neuronas en dos dimensiones para que los alumnos construyan un modelo de circuito neuronal que ocurre en distintas situaciones cotidianas tanto de actos voluntarios como de actos reflejos. La explicación del modelo se graba en vídeo. Para la evaluación de la actividad se dispone de instrumentos variados incluyendo rúbricas para la auto- y coevaluación

Intracellular <i>Salmonella</i> induces aggrephagy of host endomembranes in persistent infections

<p>Xenophagy has been studied in epithelial cells infected with <i>Salmonella enterica</i> serovar Typhimurium (<i>S</i>. Typhimurium). Distinct autophagy receptors target this pathogen to degradation after interacting with ubiquitin on the surface of cytosolic bacteria, and the phagophore- and autophagosome-associated protein MAP1LC3/LC3. Glycans exposed in damaged phagosomal membranes and diacylglycerol accumulation in the phagosomal membrane also trigger <i>S</i>. Typhimurium xenophagy. How these responses control intraphagosomal and cytosolic bacteria remains poorly understood. Here, we examined <i>S</i>. Typhimurium interaction with autophagy in fibroblasts, in which the pathogen displays limited growth and does not escape into the cytosol. Live-cell imaging microscopy revealed that <i>S</i>. Typhimurium recruits late endosomal or lysosomal compartments that evolve into a membranous aggregate connected to the phagosome. Active dynamics and integrity of the phagosomal membrane are requisite to induce such aggregates. This membranous structure increases over time to become an aggresome that engages autophagy machinery at late infection times (> 6 h postentry). The newly formed autophagosome harbors LC3 and the autophagy receptor SQSTM1/p62 but is devoid of ubiquitin and the receptor CALCOCO2/NDP52. Live-cell imaging showed that this autophagosome captures and digests within the same vacuole the aggresome and some apposed intraphagosomal bacteria. Other phagosomes move away from the aggresome and avoid destruction. Thus, host endomembrane accumulation resulting from activity of intracellular <i>S</i>. Typhimurium stimulates a novel type of aggrephagy that acts independently of ubiquitin and CALCOCO2, and destroys only a few bacteria. Such selective degradation might allow the pathogen to reduce its progeny and, as a consequence, to establish persistent infections.</p

Péptidos que contienen D-Alanina (D-Ala) o aminoalcoholes relacionados

[EN] The invention relates to peptides of general formula S, wherein the carboxyl terminus (D-alanine, D-Ala) is replaced by an analogous amino alcohol. The invention also relates to the method for synthesising same and the use of the aforementioned peptides to treat inflammatory diseases. Formula (I)[ES] La invención se refiere a péptidos de fórmula general S, donde el extremo carboxilo terminal (D-Alanina, D-Aia) es reemplazado por un amlnoalcohol análogo. La invención también se refiere a su procedimiento de síntesis y al uso de los péptidos mencionados para el tratamiento de enfermedades inflamatorias. Formula (I)[FR] L'invention concerne des peptides de formule générale S, où l'extrémité carboxyle terminale (D-alanine, D-ala) est remplacée par un amino-alcool analogue. L'invention concerne également son procédé de synthèse et l'utilisation des peptides mentionnés pour le traitement de maladies inflammatoires. Formule (I)Peer reviewedConsejo Superior de Investigaciones Científicas (España), Universidad Autónoma de MadridA1 Solicitud de patente con informe sobre el estado de la técnic