Perez-García, J. (José)

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    Molecular features of metaplastic breast carcinoma: An infrequent subtype of triple negative breast carcinoma
    (MDPI AG, 2020) Palacios, J. (José); Cortes, J. (Javier); Pérez-Mies, B. (Belen); Carretero-Barrio, I. (Irene); González-Martínez, S. (Silvia); Perez-García, J. (José); Palacios-Berraquero, M.L. (María L.)
    Metaplastic breast carcinoma (MBC) is a heterogeneous group of infrequent invasive carcinomas that display differentiation of the neoplastic epithelium towards squamous cells and/or mesenchymal-type elements. Most MBC have a triple negative phenotype and poor prognosis. Thus, MBC have worse survival rates than other invasive breast carcinomas, including other triple negative breast carcinomas (TNBC). In this study, we reviewed the molecular features of MBC, pointing out the differences among subtypes. The most frequently mutated genes in MBC were TP53 and PIK3CA. Additionally, mutations in the other genes of the PI3K/AKT pathway indicated its importance in the pathogenesis of MBC. Regarding copy number variations (CNVs), MYC was the most frequently amplified gene, and the most frequent gene loss affected the CDKN2A/CDKN2B locus. Furthermore, the pattern of mutations and CNVs of MBC differed from those reported in other TNBC. However, the molecular profile of MBC was not homogeneous among histological subtypes, being the alterations in the PI3K pathway most frequent in spindle cell carcinomas. Transcriptomic studies have demonstrated an epithelial to mesenchymal program activation and the enrichment of stemness genes in most MBC. In addition, current studies are attempting to define the immune microenvironment of these tumors. In conclusion, due to specific molecular features, MBC have a different clinical behavior from other types of TNBC, being more resistant to standard chemotherapy. For this reason, new therapeutic approaches based on tumor molecular characteristics are needed to treat MBC.
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    Comparison of six commercial serum exosome isolation methods suitable for clinical laboratories. Effect in cytokine analysis
    (2019) Alegre-Martinez, E. (Estibaliz); Rebmann, V. (Vera); Varo, N. (Nerea); Gonzalez-Hernandez, A. (Alvaro); Mateos, B. (Beatriz); Macías, M. (Mónica); Perez-García, J. (José)
    Background: Exosomes are nanovesicles released by cells that can be detected in blood. Exosomes contain several molecules, such as cytokines that have potential utility as disease biomarkers. The aim of the present work is to compare six different commercial kits suitable for the clinical laboratory in relation to the efficiency and purity of exosome isolation, and their effect in subsequent cytokines analysis. Methods: Serum exosomes were obtained from 10 volunteers using six commercial kits: exoEasy, ExoQuick, Exo-spin, ME kit, ExoQuick Plus and Exo-Flow. Exosome concentrations and size distributions were quantified by nanoparticle tracking analysis. Exosome markers CD63, CD9 and TSG101 were determined by Western blot. ApoB and albumin were measured using nephelometry. S100A9, CXCL5 and CXCL12 were measured using a Luminex assay. Results: The concentration of particles obtained between different kits varied by a factor of 100. There was no correlation in particle concentrations extracted between different kits, except between ExoQuick and Exo-Flow. The highest exosome purity was achieved with ExoQuick Plus and exoEasy, while the lowest were achieved with ME and ExoQuick. Albumin was present in all exosome extracts analyzed and ApoB in all except those extracted with Exo-Flow and ME. Cytokine detection varied depending on the purification kit used and there was no correlation in cytokine concentrations between samples obtained with different kits. Conclusions: Both the sample and the type of commercial kit used affect the efficiency and purity of exosome isolation. In addition, the exosome purification method deeply affects the capability to detect and quantify cytokines.
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    Intratumoral nanoplexed poly I:C BO-112 in combination with systemic anti–PD-1 for patients with anti–PD-1–refractory tumors
    (2020) Longo, F. (Federico); Tersago, D. (Dominique); Ponce, S. (Santiago); Jove, M. (Maria); Calles, A. (Antonio); Sempere-Ortega, C. (Cayetano); Martín-Echarri, M. (Miguel); Soria, A. (Ainara); López-Tarruella, S. (Sara); Martin-Algarra, S. (Salvador); Rodriguez-Ruiz, M.E. (María Esperanza); Aznar, M.A. (María Ángela); Jiménez-Aguilar, E. (Elisabeth); Castañon, E. (Eduardo); Marquez-Rodas, I. (Iván); Alvarez, R. (Rosa); Calvo, A. (Aitana); Quintero, M. (Marisol); Ponz-Sarvise, M. (Mariano); Gómez-Rueda, A. (Ana); Lopez-Casas, P. (Pedro); Melero, I. (Ignacio); Rubio, B. (Belén); de Miguel, E. (Enrique); Gajate, P. (Pablo); Perez-García, J. (José); Ramos-Medina, R. (Rocío)
    Intratumoral therapies, especially Toll-like receptor agonists, can trigger both the innate and adaptive immune systems. BO-112 is a nanoplexed form of polyinosinic:polycytidylic acid (poly I:C) that induces local and systemic immunotherapeutic effects in mouse models. In a multicenter phase 1 clinical trial, repeated intratumoral administrations of BO-112 induced an increase in tumor cell necrosis and apoptosis, as well as augmented immune reactivity according to gene expression profiling. The first three cohorts receiving BO-112 as a monotherapy resulted in a recommended dose of 1 mg that could be safely repeated. Two grade 3 to 4 adverse reactions in the form of reversible thrombocytopenia were reported. In a fourth cohort of 28 patients with tumors that had primary resistance to anti–programmed cell death protein–1 (PD-1), the combination of intratumoral BO-112 with nivolumab or pembrolizumab was also well tolerated, and 3 patients (2 with melanoma and 1 with renal cell carcinoma) achieved partial responses, with 10 more patients having stable disease at 8 to 12 weeks. Thus, local BO-112 combined with a systemic anti–PD-1 agent might be a strategy to revert anti–PD-1 resistance