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Spliceosomic dysregulation unveils NOVA1 as a candidate actionable therapeutic target in pancreatic neuroendocrine tumors

  • Sergio Pedraza-Arevalo
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Emilia Alors-Pérez
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Ricardo Blázquez-Encinas
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Aura D. Herrera-Martínez
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Endocrinology and Nutrition Service, Reina Sofia University Hospital, Córdoba, Spain
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  • Juan M. Jiménez-Vacas
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Antonio C. Fuentes-Fayos
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Óscar Reyes
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Computer Sciences, University of Córdoba, Córdoba, Spain
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  • Sebastián Ventura
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Computer Sciences, University of Córdoba, Córdoba, Spain
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  • Rafael Sánchez-Sánchez
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Pathology Service, Reina Sofia University Hospital, Córdoba, Spain
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  • Rosa Ortega-Salas
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Pathology Service, Reina Sofia University Hospital, Córdoba, Spain
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  • Raquel Serrano-Blanch
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Medical Oncology Service, Reina Sofia University Hospital, Córdoba, Spain
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  • María A. Gálvez-Moreno
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Endocrinology and Nutrition Service, Reina Sofia University Hospital, Córdoba, Spain
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  • Manuel D. Gahete
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
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  • Author Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Alejandro Ibáñez-Costa
    Correspondence
    Reprint requests: J.P. Castaño, Alejandro Ibáñez-Costa, and Raúl M. Luque Avenida Menéndez Pidal s/n, Edificio IMIBIC, 14004, Córdoba, Spain.
    Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
    Search for articles by this author
  • Author Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Raúl M. Luque
    Correspondence
    Reprint requests: J.P. Castaño, Alejandro Ibáñez-Costa, and Raúl M. Luque Avenida Menéndez Pidal s/n, Edificio IMIBIC, 14004, Córdoba, Spain.
    Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
    Search for articles by this author
  • Author Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Justo P. Castaño
    Correspondence
    Reprint requests: J.P. Castaño, Alejandro Ibáñez-Costa, and Raúl M. Luque Avenida Menéndez Pidal s/n, Edificio IMIBIC, 14004, Córdoba, Spain.
    Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.
    Affiliations
    Maimonides Biomedical Research Institute of Córdoba (IMIBIC), Córdoba, Spain

    Department of Cell Biology, Physiology, and Immunology, University of Córdoba, Córdoba, Spain

    CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), Córdoba, Spain

    Reina Sofia University Hospital, Córdoba, Spain
    Search for articles by this author
  • Author Footnotes
    # A.I.C., R. M. L. and J. P. C. contributed equally to this study and are co-senior authors.

      Abstract

      Dysregulation of the splicing machinery is emerging as a hallmark in cancer due to its association with multiple dysfunctions in tumor cells. Inappropriate function of this machinery can generate tumor-driving splicing variants and trigger oncogenic actions. However, its role in pancreatic neuroendocrine tumors (PanNETs) is poorly defined. In this study we aimed to characterize the expression pattern of a set of splicing machinery components in PanNETs, and their relationship with aggressiveness features. A qPCR-based array was first deployed to determine the expression levels of components of the major (n = 13) and minor spliceosome (n = 4) and associated splicing factors (n = 27), using a microfluidic technology in 20 PanNETs and non-tumoral adjacent samples. Subsequently, in vivo and in vitro models were applied to explore the pathophysiological role of NOVA1. Expression analysis revealed that a substantial proportion of splicing machinery components was altered in tumors. Notably, key splicing factors were overexpressed in PanNETs samples, wherein their levels correlated with clinical and malignancy features. Using in vivo and in vitro assays, we demonstrate that one of those altered factors, NOVA1, is tightly related to cell proliferation, alters pivotal signaling pathways and interferes with responsiveness to drug treatment in PanNETs, suggesting a role for this factor in the aggressiveness of these tumors and its suitability as therapeutic target. Altogether, our results unveil a severe alteration of the splicing machinery in PanNETs and identify the putative relevance of NOVA1 in tumor development/progression, which could provide novel avenues to develop diagnostic biomarkers and therapeutic tools for this pathology.

      Abbreviations:

      CgA (chromogranin A), FBS (fetal bovine serum), FFPE (formalin-fixed paraffin-embedded), IHC (immunohistochemistry), MAPK (Mitogen-activated protein kinase), mTOR (mammalian target of rapamycin), NETs (neuroendocrine tumors), PanNETs (Pancreatic neuroendocrine tumors), PI3K (phosphatidylinositol 3-kinase), qPCR (quantitative polymerase chain reaction RNA), ROC (receiver operating characteristic), snRNAs (small nuclear RNA), siRNA (small interference)
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