Person: García-Fojeda García-Valdecasas, María Belén
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First Name
María Belén
Last Name
García-Fojeda García-Valdecasas
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Bioquímica y Biología Molecular
Area
Bioquímica y Biología Molecular
Identifiers
12 results
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Now showing 1 - 10 of 12
Item Apoptosis, Toll-like, RIG-I-like and NOD-like Receptors Are Pathways Jointly Induced by Diverse Respiratory Bacterial and Viral Pathogens(Frontiers in Immunology, 2017) Martínez, Isidoro ; Oliveros, Juan; Cuesta, Isabel; Barrera, Jorge; Ausina, Vicente; Casals Carro, María Cristina; Lorenzo, Alba de ; García, Ernesto; García-Fojeda García-Valdecasas, María Belén; Garmendia, Junkal ; González-Nicolau, Mar; Lacoma, Alicia; Menéndez, Margarita; Moranta, David ; Nieto, Amelia ; Ortín, Juan; Pérez-González, Alicia ; Prat, Cristina ; Ramos-Sevillano, Elisa ; Regueiro, Verónica; Rodriguez-Frandsen, Ariel ; Solís, Dolores ; Yuste, José ; Bengoechea, José ; Melero, JoséLower respiratory tract infections are among the top five leading causes of human death. Fighting these infections is therefore a world health priority. Searching for induced alterations in host gene expression shared by several relevant respiratory pathogens represents an alternative to identify new targets for wide-range host-oriented therapeutics. With this aim, alveolar macrophages were independently infected with three unrelated bacterial (Streptococcus pneumoniae, Klebsiella pneumoniae, and Staphylococcus aureus) and two dissimilar viral (respiratory syncytial virus and influenza A virus) respiratory pathogens, all of them highly relevant for human health. Cells were also activated with bacterial lipopolysaccharide (LPS) as a prototypical pathogen-associated molecular pattern. Patterns of differentially expressed cellular genes shared by the indicated pathogens were searched by microarray analysis. Most of the commonly up-regulated host genes were related to the innate immune response and/or apoptosis, with Toll-like, RIG-I-like and NOD-like receptors among the top 10 signaling pathways with over-expressed genes. These results identify new potential broad-spectrum targets to fight the important human infections caused by the bacteria and viruses studied here.- Project number: 163
Item The Phantom Menace: Cómo salvar el mundo de una pandemia mediante Ingeniería Genética cooperativa(2021) Navarro LLorens, Juana María; Baldanta Callejo, Sara; Bénitez Prian, Mario; Blázquez Ortiz, Cristina; Bruñen Alfaro, Francisco; Cañadas Benito, Olga; Chinarro Sánchez, Adrián; García de la Camacha Selgas, Nuria; García-Fojeda García-Valdecasas, María Belén; Guevara Acosta, Flor Govinda; Leaño Hinojosa, Ariana; López Conejo, Maria Teresa; Lorente Pérez, Maria del Mar; Nogués Vera, Laura; Penalba Iglesias, Diana; Raisman, Andrea; Ranz Valdecasa, Maria Regina; Ruiz Ortega, Marta; Sánchez-Escalonilla Relea, Jose luis; Velasco Díez, Guillermo; Sánchez Torralba, AntonioEl año pasado se llevó a cabo un proyecto de innovación (PIMCD-2019 174) basado en el modelo pedagógico de la clase invertida o “Flipped Classroom” y en el juego de mesa PANDEMIC (ASMODEE IBÉRICA). Brevemente, este proyecto consistió en una propuesta didáctica sobre los contenidos de FIGG en el que tomando como partida un entorno lúdico, los alumnos de FIGG en equipos preparaban contenidos y reforzaban conocimientos adquiridos en clase. Para llevarlo a cabo, a los alumnos se les planteaba una hipotética situación en la que cuatro enfermedades mortales aparecen en la tierra. Los alumnos divididos en equipos de 5 personas, deben cooperar desarrollando una serie de acciones que les permitan ir conociendo a qué patógeno se enfrentan y desarrollar herramientas que les permita controlar la epidemia dentro de un tiempo dado. Así cada equipo debe frenar el avance de las infecciones y a la vez encontrar una cura para la misma. Curiosamente, todo este escenario fue planteado antes de que la crisis del COVID-19 impactara en nuestra sociedad. En cualquier caso, la situación vivida en los últimos meses ha motivado a los estudiantes de FIGG por lo que en su mayor parte han participado en su desarrollo de manera entusiasta. En este nuevo proyecto, hemos tomado como base el proyecto del curso pasado, pero introduciendo algunos elementos innovadores. Entre estas innovaciones destaca la incorporación de dos alumnos por grupo de FIGG que ya lo hubieran realizado en la edición pasada por cada uno de los grupos de FIGG como mentores. Item Laforin, the dual-phosphatase responsible for Lafora disease, interacts with R5 (PTG), a regulatory subunit of protein phosphatase-1 that enhances glycogen accumulation(Human Molecular Genetics, 2003) Fernández-Sánchez, M. E.; Criado-García, O.; Heath, K. E.; García-Fojeda García-Valdecasas, María Belén; Medraño-Fernández, Iria; Gómez-Garre, Pilar; Sanz, Pascual; Serratosa, José María; Rodríguez de Córdoba, SantiagoProgressive myoclonus epilepsy of Lafora type (LD, MIM 254780) is a fatal autosomal recessive disorder characterized by the presence of progressive neurological deterioration, myoclonus, epilepsy and polyglucosan intracellular inclusion bodies, called Lafora bodies. Lafora bodies resemble glycogen with reduced branching, suggesting an alteration in glycogen metabolism. Linkage analysis and homozygosity mapping localized EPM2A, a major gene for LD, to chromosome 6q24. EPM2A encodes a protein of 331 amino acids (named laforin) with two domains, a dual-specificity phosphatase domain and a carbohydrate binding domain. Here we show that, in addition, laforin interacts with itself and with the glycogen targeting regulatory subunit R5 of protein phosphatase 1 (PP1). R5 is the human homolog of the murine Protein Targeting to Glycogen, a protein that also acts as a molecular scaffold assembling PP1 with its substrate, glycogen synthase, at the intracellular glycogen particles. The laforin–R5 interaction was confirmed by pull-down and co-localization experiments. Full-length laforin is required for the interaction. However, a minimal central region of R5 (amino acids 116–238), including the binding sites for glycogen and for glycogen synthase, is sufficient to interact with laforin. Point-mutagenesis of the glycogen synthase-binding site completely blocked the interaction with laforin. The majority of the EPM2A missense mutations found in LD patients result in lack of phosphatase activity, absence of binding to glycogen and lack of interaction with R5. Interestingly, we have found that the LD-associated EPM2A missense mutation G240S has no effect on the phosphatase or glycogen binding activities of laforin but disrupts the interaction with R5, suggesting that binding to R5 is critical for the laforin function. These results place laforin in the context of a multiprotein complex associated with intracellular glycogen particles, reinforcing the concept that laforin is involved in the regulation of glycogen metabolism.- Project number: PIMCD327/23-24
Item El poder de la nanotecnología lipídica: aplicaciones biotecnológicas de la asignatura Estructura de Membranas Biológicas(2024) Cañadas Benito, Olga; Ballesteros Aparicio, Sara; Cruz Rodríguez, Antonio; Espada Espinar, Adrián; Fuertes Vázquez, Lucía; García-Fojeda García-Valdecasas, María Belén; Gutiérrez Ramos, Irene Item Mechanism suppressing glycogen synthesis in neurons and its demise in progressive myoclonus epilepsy(Nature Neuroscience, 2007) Vilchez, David; Ros, Susana; Cifuentes, Daniel; Pujadas, Lluís; Vallès, Jordi; García-Fojeda García-Valdecasas, María Belén; Criado-García, Olga; Fernández-Sánchez, Elena; Medraño-Fernández, Iria; Domínguez, Jorge; García-Rocha, Mar; Soriano, Eduardo; Rodríguez de Córdoba, Santiago; Guinovart, JoanGlycogen synthesis is normally absent in neurons. However, inclusion bodies resembling abnormal glycogen accumulate in several neurological diseases, particularly in progressive myoclonus epilepsy or Lafora disease. We show here that mouse neurons have the enzymatic machinery for synthesizing glycogen, but that it is suppressed by retention of muscle glycogen synthase (MGS) in the phosphorylated, inactive state. This suppression was further ensured by a complex of laforin and malin, which are the two proteins whose mutations cause Lafora disease. The laforin-malin complex caused proteasome-dependent degradation both of the adaptor protein targeting to glycogen, PTG, which brings protein phosphatase 1 to MGS for activation, and of MGS itself. Enforced expression of PTG led to glycogen deposition in neurons and caused apoptosis. Therefore, the malin-laforin complex ensures a blockade of neuronal glycogen synthesis even under intense glycogenic conditions. Here we explain the formation of polyglucosan inclusions in Lafora disease by demonstrating a crucial role for laforin and malin in glycogen synthesis.Item Surfactant Protein A Prevents IFN-γ/IFN-γ Receptor Interaction and Attenuates Classical Activation of Human Alveolar Macrophages(Journal of Immunology, 2016) Minutti, Carlos; García-Fojeda García-Valdecasas, María Belén; Sáenz, Alejandra ; Casas-Engel, Mateo de las ; Guillamat-Prats, Raquel ; Lorenzo, Alba de ; Serrano-Mollar, Anna ; Corbí, Ängel; Casals Carro, María CristinaLung surfactant protein A (SP-A) plays an important function in modulating inflammation in the lung. However, the exact role of SP-A and the mechanism by which SP-A affects IFN-γ–induced activation of alveolar macrophages (aMϕs) remains unknown. To address these questions, we studied the effect of human SP-A on rat and human aMϕs stimulated with IFN-γ, LPS, and combinations thereof and measured the induction of proinflammatory mediators as well as SP-A’s ability to bind to IFN-γ or IFN-γR1. We found that SP-A inhibited (IFN-γ + LPS)–induced TNF-α, iNOS, and CXCL10 production by rat aMϕs. When rat macrophages were stimulated with LPS and IFN-γ separately, SP-A inhibited both LPS-induced signaling and IFN-γ–elicited STAT1 phosphorylation. SP-A also decreased TNF-α and CXCL10 secretion by ex vivo–cultured human aMϕs and M-CSF–derived macrophages stimulated by either LPS or IFN-γ or both. Hence, SP-A inhibited upregulation of IFN-γ–inducible genes (CXCL10, RARRES3, and ETV7) as well as STAT1 phosphorylation in human M-CSF–derived macrophages. In addition, we found that SP-A bound to human IFN-γ (KD = 11 ± 0.5 nM) in a Ca2+-dependent manner and prevented IFN-γ interaction with IFN-γR1 on human aMϕs. We conclude that SP-A inhibition of (IFN-γ + LPS) stimulation is due to SP-A attenuation of both inflammatory agents and that the binding of SP-A to IFN-γ abrogates IFN-γ effects on human macrophages, suppressing their classical activation and subsequent inflammatory response.Item The collectin SP-A and its trimeric recombinant fragment protect alveolar epithelial cells from the cytotoxic and proinflammatory effects of human cathelicidin in vitro(Frontiers in Immunology, 2022) De Tapia Hernández, Lidia Consuelo; García-Fojeda García-Valdecasas, María Belén; Kronqvist, Nina; Johansson, Jan; Casals Carro, María CristinaHuman cathelicidin (LL-37) is a defense peptide with antimicrobial activity against various pathogens. However, LL-37 can also trigger tissue injury by binding to host cell membranes. The cytotoxic effects of LL-37 may be especially relevant in chronic respiratory diseases characterized by increased LL-37. The aim of this study was to investigate whether the human collectin SP-A and a trimeric recombinant fragment thereof (rfhSP-A) can regulate the activities of LL-37. To this end, we studied the interaction of LL-37 with SP-A and rfhSP-A by intrinsic fluorescence, dynamic light scattering, and circular dichroism, as well as the effects of these proteins on the antimicrobial and cytotoxic activities of LL-37. Both SP-A and rfhSP-A bound LL-37 with high affinity at physiological ionic strength ( 0.45 ± 0.01 nM for SP-A and 1.22 ± 0.7 nM for rfhSP-A). Such interactions result in the reduction of LL-37-induced cell permeability and IL-8 release in human pneumocytes, mediated by P2X7 channels. Binding of LL-37 to SP-A did not modify the properties of SP-A or the antibacterial activity of LL-37 against respiratory pathogens (Pseudomonas aeruginosa, and nontypeable Haemophilus influenzae). SP-A/LL-37 complexes showed a greater ability to aggregate LPS vesicles than LL-37, which reduces endotoxin bioactivity. These results reveal the protective role of native SP-A in controlling LL-37 activities and suggest a potential therapeutic effect of rfhSP-A in reducing the cytotoxic and inflammatory actions of LL-37, without affecting its microbicidal activity against Gram-negative pathogens.Item Airway allergy causes alveolar macrophage death, profound alveolar disorganization and surfactant dysfunction(Frontiers in Immunology, 2023) Feo-Lucas, Lidia; Godio, Cristina; Minguito de la Escalera, María ; Alvarez-Ladrón, Natalia ; Villarrubia, Laura; Vega-Pérez, Adrián ; González-Cintado, Leticia ; Domínguez-Andrés, Jorge; García-Fojeda García-Valdecasas, María Belén; Montero Fernández, Carlos; Casals Carro, María Cristina; Autilio, Chiara; Pérez Gil, Jesús; Crainiciuc, Georgiana ; Hidalgo, Andrés ; López-Bravo, María ; Fernández-Ardavín Castro, CarlosRespiratory disorders caused by allergy have been associated to bronchiolar inflammation leading to life-threatening airway narrowing. However, whether airway allergy causes alveolar dysfunction contributing to the pathology of allergic asthma remains unaddressed. To explore whether airway allergy causes alveolar dysfunction that might contribute to the pathology of allergic asthma, alveolar structural and functional alterations were analyzed during house dust mite (HDM)-induced airway allergy in mice, by flow cytometry, light and electron microscopy, monocyte transfer experiments, assessment of intra-alveolarly-located cells, analysis of alveolar macrophage regeneration in Cx3cr1cre:R26-yfp chimeras, analysis of surfactant-associated proteins, and study of lung surfactant biophysical properties by captive bubble surfactometry. Our results demonstrate that HDM-induced airway allergic reactions caused severe alveolar dysfunction, leading to alveolar macrophage death, pneumocyte hypertrophy and surfactant dysfunction. SP-B/C proteins were reduced in allergic lung surfactant, that displayed a reduced efficiency to form surface-active films, increasing the risk of atelectasis. Original alveolar macrophages were replaced by monocyte-derived alveolar macrophages, that persisted at least two months after the resolution of allergy. Monocyte to alveolar macrophage transition occurred through an intermediate stage of pre-alveolar macrophage and was paralleled with translocation into the alveolar space, Siglec-F upregulation, and downregulation of CX3CR1. These data support that the severe respiratory disorders caused by asthmatic reactions not only result from bronchiolar inflammation, but additionally from alveolar dysfunction compromising an efficient gas exchange.Item Local amplifiers of IL-4Rα–mediated macrophage activation promote repair in lung and liver(Science, 2017) Minutti, Carlos; Jackson-Jones, Lucy; García-Fojeda García-Valdecasas, María Belén; Knipper, Johanna ; Sutherland, Tara; Logan, Nicola; Ringqvist, Emma; Guillamat-Prats, Raquel; Ferenbach, David; Artigas, Antonio; Stamme, Cordula ; Chroneos, Zissis ; Zaiss, Dietmar ; Casals Carro, María Cristina; Allen, JudithThe type 2 immune response controls helminth infection and maintains tissue homeostasis but can lead to allergy and fibrosis if not adequately regulated. We have discovered local tissue-specific amplifiers of type 2-mediated macrophage activation. In the lung, surfactant protein A (SP-A) enhanced interleukin-4 (IL-4)-dependent macrophage proliferation and activation, accelerating parasite clearance and reducing pulmonary injury after infection with a lung-migrating helminth. In the peritoneal cavity and liver, C1q enhancement of type 2 macrophage activation was required for liver repair after bacterial infection, but resulted in fibrosis after peritoneal dialysis. IL-4 drives production of these structurally related defense collagens, SP-A and C1q, and the expression of their receptor, myosin 18A. These findings reveal the existence within different tissues of an amplification system needed for local type 2 responses.Item Folding and Intramembraneous BRICHOS Binding of the Prosurfactant Protein C Transmembrane Segment(Journal of Biological Chemistry, 2015) Sáenz, Alejandra ; Presto. Jenny ; Lara, Patricia ; Akinyi-Oloo, Laura ; García-Fojeda García-Valdecasas, María Belén; Nilsson, IngMarie; Johansson, Jan ; Casals Carro, María CristinaSurfactant protein C (SP-C) is a novel amyloid protein found in the lung tissue of patients suffering from interstitial lung disease (ILD) due to mutations in the gene of the precursor protein pro-SP-C. SP-C is a small α-helical hydrophobic protein with an unusually high content of valine residues. SP-C is prone to convert into β-sheet aggregates, forming amyloid fibrils. Nature's way of solving this folding problem is to include a BRICHOS domain in pro-SP-C, which functions as a chaperone for SP-C during biosynthesis. Mutations in the pro-SP-C BRICHOS domain or linker region lead to amyloid formation of the SP-C protein and ILD. In this study, we used an in vitro transcription/translation system to study translocon-mediated folding of the WT pro-SP-C poly-Val and a designed poly-Leu transmembrane (TM) segment in the endoplasmic reticulum (ER) membrane. Furthermore, to understand how the pro-SP-C BRICHOS domain present in the ER lumen can interact with the TM segment of pro-SP-C, we studied the membrane insertion properties of the recombinant form of the pro-SP-C BRICHOS domain and two ILD-associated mutants. The results show that the co-translational folding of the WT pro-SP-C TM segment is inefficient, that the BRICHOS domain inserts into superficial parts of fluid membranes, and that BRICHOS membrane insertion is promoted by poly-Val peptides present in the membrane. In contrast, one BRICHOS and one non-BRICHOS ILD-associated mutant could not insert into membranes. These findings support a chaperone function of the BRICHOS domain, possibly together with the linker region, during pro-SP-C biosynthesis in the ER.