Examinando por Autor "Vallejo Illarramendi, Ainara"

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    Dysregulated FOXO1 activity drives skeletal muscle intrinsic dysfunction in amyotrophic lateral sclerosis
    (Springer Science and Business Media Deutschland GmbH, 2024-09-16) Zufiría García, Mónica; Pikatza-Menoio, Oihane; Garciandia Arcelus, Maddi; Bengoetxea Bausela, Xabier; Jiménez Zúñiga, Andrés; Elicegui, Amaia; Levchuk, María; Arnold García, Olatz; Ondaro Ezkurra, Jon; Iruzubieta Agudo, Pablo; Rodríguez Gómez, Laura; Fernández Pelayo, Uxoa; Muñoz Oreja, Mikel; Aiastui Pujana, Ana; García Verdugo, Jose Manuel; Herranz Pérez, Vicente; Zulaica, Miren; Poza Aldea, Juan José; Ruiz Onandi, Rebeca; Fernández Torrón, Roberto; Espinal Valencia, Juan Bautista; Bonilla Zagala, Mario; Lersundi Artamendi, Ana; Fernández-Eulate, Gorka; Riancho Zarrabeitia, Javier; Vallejo Illarramendi, Ainara; Holt, Ian James; Sáenz, Amets; Malfatti, Edoardo; Duguez, Stéphanie; Blázquez García, Lorea; López de Munain Arregui, Adolfo; Gereñu Lopetegi, Gorka; Gil Bea, Francisco Javier; Alonso-Martin, Sonia
    Amyotrophic Lateral Sclerosis (ALS) is a multisystemic neurodegenerative disorder, with accumulating evidence indicating metabolic disruptions in the skeletal muscle preceding disease symptoms, rather than them manifesting as a secondary consequence of motor neuron (MN) degeneration. Hence, energy homeostasis is deeply implicated in the complex physiopathology of ALS and skeletal muscle has emerged as a key therapeutic target. Here, we describe intrinsic abnormalities in ALS skeletal muscle, both in patient-derived muscle cells and in muscle cell lines with genetic knockdown of genes related to familial ALS, such as TARDBP (TDP-43) and FUS. We found a functional impairment of myogenesis that parallels defects of glucose oxidation in ALS muscle cells. We identified FOXO1 transcription factor as a key mediator of these metabolic and functional features in ALS muscle, via gene expression profiling and biochemical surveys in TDP-43 and FUS-silenced muscle progenitors. Strikingly, inhibition of FOXO1 mitigated the impaired myogenesis in both the genetically modified and the primary ALS myoblasts. In addition, specific in vivo conditional knockdown of TDP-43 or FUS orthologs (TBPH or caz) in Drosophila muscle precursor cells resulted in decreased innervation and profound dysfunction of motor nerve terminals and neuromuscular synapses, accompanied by motor abnormalities and reduced lifespan. Remarkably, these phenotypes were partially corrected by foxo inhibition, bolstering the potential pharmacological management of muscle intrinsic abnormalities associated with ALS. The findings demonstrate an intrinsic muscle dysfunction in ALS, which can be modulated by targeting FOXO factors, paving the way for novel therapeutic approaches that focus on the skeletal muscle as complementary target tissue.
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    A founder variant in the RYR1 gene is associated with hyperCKemia, myalgia and muscle cramps
    (John Wiley and Sons Inc, 2025-01) Segarra Casas, Alba; Iruzubieta Agudo, Pablo; Kapetanovic García, Solange; Hernández Laín, A.; Jericó Pascual, Ivonne; Fernández Torrón, Roberto; Maneiro, Miren; Marco Moreno, Pablo; Zelaya Huerta, María Victoria; Rodríguez Santiago, Benjamín; Calafell Majó, Francesc; Töpf, Ana; Straub, Volker; Vallejo Illarramendi, Ainara; López de Munain Arregui, Adolfo; Gallano Petit, María Pía; González Quereda, Lidia
    Background and purpose: Pathogenic variants in the RYR1 gene have been associated with a variety of conditions, ranging from congenital myopathy to adult manifestations. Our aim was to characterize the p.Leu2286Val variant in 17 Basque patients, to accurately determine its correlation with clinical features and to explore the possible founder effect of the variant. Methods: Families harbouring the p.Leu2286 RYR1 variant underwent a detailed clinical evaluation, including muscle magnetic resonance imaging, electromyography and muscle biopsy. Haplotypes were analysed in available patients and their relatives. Results: Individuals carrying the p.Leu2286Val shared a common haplotype, suggesting a founder event in the Basque Country population. The most prevalent features were exertional myalgia, high creatine kinase (CK) levels, cramps and muscle hypertrophy. None of the patients carrying only the p.Leu2286Val showed progression to severe muscle weakness and muscle magnetic resonance imaging showed a heterogeneous muscle involvement. Muscle biopsy revealed non-specific findings in two patients and features associated with central core disease in one patient carrying only the p.Leu2286Val and two patients harbouring an additional RYR1 variant. Three individuals carrying an in trans RYR1 variant presented with an earlier onset and more severe phenotype. Conclusion: Here, it is shown that the dominantly inherited p.Leu2286Val RYR1 founder variant is associated with a milder phenotype of exercise intolerance, myalgia and hyperCKemia.
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    A novel class of FKBP12 ligands rescues premature aging phenotypes associated with myotonic dystrophy type 1
    (Multidisciplinary Digital Publishing Institute (MDPI), 2024-12-01) García Puga, Mikel; Gereñu Lopetegi, Gorka; Bargiela Schönbrunn, Ariadna; Espinosa Espinosa, Jorge; Mosqueira Martín, Laura; Sagartzazu Aizpurua, Maialen; Aizpurua Iparraguirre, Jesus Mari; Vallejo Illarramendi, Ainara; Artero Allepuz, Rubén; López de Munain Arregui, Adolfo; Matheu Fernández, Ander
    Background: Myotonic dystrophy type 1 (DM1) is an autosomal dominant disorder clinically characterized by progressive muscular weakness and multisystem degeneration, which correlates with the size of CTG expansion and MBLN decrease. These changes induce a calcium and redox homeostasis imbalance in several models that recapitulate the features of premature tissue aging. In this study, we characterized the impact of a new family of FKBP12 ligands (generically named MPs or MP compounds) designed to stabilize FKBP12 binding to the ryanodine receptors and normalize calcium dysregulation under oxidative stress. Methods: Human primary fibroblasts from DM1 patients and control donors, treated with MP compounds or not, were used for functional studies of cell viability, proliferation, and metabolism. The gene expression profile in treated cells was determined using RNA sequencing. The impact of MP compounds in vivo was evaluated in a Drosophila model of the disease using locomotor activity and longevity studies. Results: The treatment with different MP compounds reversed oxidative stress and impaired cell viability and proliferation, mitochondrial activity, and metabolic defects in DM1-derived primary fibroblasts. RNA sequencing analysis confirmed the restoration of molecular pathways related to calcium and redox homeostasis and additional pathways, including the cell cycle and metabolism. This analysis also revealed the rescue of alternative splicing events in DM1 fibroblasts treated with MP compounds. Importantly, treatment with MP compounds significantly extended the lifespan and improved the locomotor activity of a Drosophila model of the DM1 disease, and restored molecular defects characteristic of the disease in vivo. Conclusions: Our results revealed that MP compounds rescue multiple premature aging phenotypes described in DM1 models and decipher the benefits of this new family of compounds in the pre-clinical setting of DM1.
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    Pharmacokinetic evaluation of new drugs using a multi-labelling approach and PET imaging: application to a drug candidate with potential application in neuromuscular disorders
    (MDPI, 2023-01-18) Passannante, Rossana; Gómez Vallejo, Vanessa; Sagartzazu Aizpurua, Maialen; Vignau Arsuaga, Laura; Marco Moreno, Pablo; Aldanondo Aristizabal, Garazi ; Vallejo Illarramendi, Ainara; Aguiar Fernández, Pablo; Cossío, Unai; Martín, Abraham; Bergare, Jonas; Kingston, Lee; Elmore, Charles S.; Morcillo Alonso, Miguel Ángel ; Ferrón, Pablo; Aizpurua Iparraguirre, Jesus Mari ; Llop, Jordi
    Background and objective: The determination of pharmacokinetic properties of new chemical entities is a key step in the process of drug development. Positron emission tomography (PET) is an ideal technique to obtain both biodistribution and pharmacokinetic parameters of new compounds over a wide range of chemical modalities. Here, we use a multi-radionuclide/multi-position labelling approach to investigate distribution, elimination, and metabolism of a triazole-based FKBP12 ligand (AHK2) with potential application in neuromuscular disorders. Methods: Target engagement and stabilizing capacity of the drug candidate (AHK2) towards FKBP12-RyR was evaluated using competitive ligand binding and proximity ligation assays, respectively. Subsequently, AHK2 was labelled either with the positron emitter carbon-11 (11C) via 11C-methylation to yield both [11C]AHK2.1 and [11C]AHK2.2, or by palladium-catalysed reduction of the corresponding 5-iodotriazole derivative using 3H gas to yield [3H]AHK2. Metabolism was first investigated in vitro using liver microsomes. PET imaging studies in rats after intravenous (IV) administration at different doses (1 µg/Kg and 5 mg/Kg) were combined with determination of arterial blood time-activity curves (TACs) and analysis of plasma samples by high performance liquid chromatography (HPLC) to quantify radioactive metabolites. Arterial TACs were obtained in continuous mode by using an in-house developed system that enables extracorporeal blood circulation and continuous measurement of radioactivity in the blood. Pharmacokinetic parameters were determined by non-compartmental modelling of the TACs. Results: In vitro studies indicate that AHK2 binds to FKBP12 at the rapamycin-binding pocket, presenting activity as a FKBP12/RyR stabilizer. [11C]AHK2.1, [11C]AHK2.2 and [3H]AHK2 could be obtained in overall non-decay corrected radiochemical yields of 14 ± 2%, 15 ± 2% and 0.05%, respectively. Molar activities were 60–110 GBq/µmol, 68–122 GBq/µmol and 0.4–0.5 GBq/μmol, respectively. In vitro results showed that oxidation of the thioether group into sulfoxide, demethylation of the CH3O-Ar residue and demethylation of –N(CH3)2 were the main metabolic pathways. Fast metabolism was observed in vivo. Pharmacokinetic parameters obtained from metabolite-corrected arterial blood TACs showed a short half-life (12.6 ± 3.3 min). Dynamic PET imaging showed elimination via urine when [11C]AHK2.2 was administered, probably reflecting the biodistribution of [11C]methanol as the major metabolite. Contrarily, accumulation in the gastrointestinal track was observed after administration of [11C]AKH2.1. Conclusions: AHK2 binds to FKBP12 at the rapamycin-binding pocket, presenting activity as a FKBP12/RyR stabilizer. Studies performed with the 3H- and 11C-labelled FKBP12/RyR stabilizer AHK2 confirm fast blood clearance, linear pharmacokinetics and rapid metabolism involving oxidation of the sulfide and amine moieties and oxidative demethylation of the CH3-O-Ar and tertiary amine groups as the main pathways. PET studies suggest that knowledge about metabolic pathways is paramount to interpret images.