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Identification of Homoharringtonine as a potent inhibitor of glioblastoma cell proliferation and migration

  • Elena Porcù
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Author Footnotes
    # Current address: Arnie Charbonneau Cancer Institute, Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3280 Hospital Drive NW - 2AA-07Calgary, AB, Canada.
    Francesca Maule
    Footnotes
    # Current address: Arnie Charbonneau Cancer Institute, Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3280 Hospital Drive NW - 2AA-07Calgary, AB, Canada.
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy
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  • Lorenzo Manfreda
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Elena Mariotto
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Silvia Bresolin
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Alice Cani
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Roberta Bortolozzi
    Affiliations
    Pediatric Research Institute, Padova, Italy
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  • Author Footnotes
    $ Current address: Neurosurgery Clinic, Department of Neuroscience, Psychology, Pharmacology and Child Health, Careggi University Hospital and University of Florence, Viale Pieraccini, 6 - 50139, Florence, Italy.
    Alessandro Della Puppa
    Footnotes
    $ Current address: Neurosurgery Clinic, Department of Neuroscience, Psychology, Pharmacology and Child Health, Careggi University Hospital and University of Florence, Viale Pieraccini, 6 - 50139, Florence, Italy.
    Affiliations
    Neurosurgery Unit, University Hospital of Padova, Padova, Italy
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  • Diana Corallo
    Affiliations
    Laboratory of Target Discovery and Biology of Neuroblastoma, Pediatric Research Institute, Padova, Italy
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  • Giampietro Viola
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Elena Rampazzo
    Correspondence
    Reprint requests: Elena Rampazzo, Department of Women's and Children's Health - University of Padova, Via Giustiniani, 3 - 35128 Padova, Italy.
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Luca Persano
    Affiliations
    Department of Women and Children's Health, University of Padova, Padova, Italy

    Pediatric Research Institute, Padova, Italy
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  • Author Footnotes
    # Current address: Arnie Charbonneau Cancer Institute, Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3280 Hospital Drive NW - 2AA-07Calgary, AB, Canada.
    $ Current address: Neurosurgery Clinic, Department of Neuroscience, Psychology, Pharmacology and Child Health, Careggi University Hospital and University of Florence, Viale Pieraccini, 6 - 50139, Florence, Italy.

      Abstract

      We previously demonstrated that Annexin A2 (ANXA2) is a pivotal mediator of the pro-oncogenic features displayed by glioblastoma (GBM) tumors, the deadliest adult brain malignancies, being involved in cell stemness, proliferation and invasion, thus negatively impacting patient prognosis. Based on these results, we hypothesized that compounds able to revert ANXA2-dependent transcriptional features could be exploited as reliable treatments to inhibit GBM cell aggressiveness by hampering their proliferative and migratory potential. Transcriptional signatures obtained by the modulation of ANXA2 activity/levels were functionally mapped through the QUADrATiC bioinformatic tool for compound identification. Selected compounds were screened by cell proliferation and migration assays in primary GBM cells, and we identified Homoharringtonine (HHT) as a potent inhibitor of GBM cell motility and proliferation, without affecting their viability. A further molecular characterization of the effects displayed by HHT, confirmed its ability to inhibit a transcriptional program involved in cell migration and invasion. Moreover, we demonstrated that the multiple antitumoral effects displayed by HHT are correlated to the inhibition of a platelet derived growth factor receptor α (PDGFRα)-dependent intracellular signaling through the impairment of Signal transducer and activator of transcription 3 (STAT3) and Ras homolog family member A (RhoA) axes. Our results demonstrate that HHT may act as a potent inhibitor of cancer cell proliferation and invasion in GBM, by hampering multiple PDGFRα-dependent oncogenic signals transduced through the STAT3 and RhoA intracellular components, finally suggesting its potential transferability for achieving an effective impairment of peculiar GBM hallmarks.

      Abbreviations:

      5-ALA (5-aminolevulinic acid), ANXA2 (Annexin A2), AZA (5′-Azacytidine), bFGF (basic Fibroblast Growth Factor), BOR (Bortezomib), CML (Chronic Myeloid Leukemia), DEGs (Differentially Expressed Genes), DIG (Digitoxin), ECM (Extracellular Matrix), EGF (Epidermal Growth Factor), EMT (Epithelial to Mesenchymal Transition), FA (Focal Adhesion), FBS (Foetal Bovine Serum), FDA (Food and Drug Administration), GBM (Glioblastoma), GEP (Gene Expression Profile), GI50 (Growth Inhibition 50), gw (gestational weeks), HHT (Homoharringtonine), hpi (hours post-injection), MFI (Mean Fluorescence Intensity), NIC (Niclosamide), OUA (Ouabain), PDGFRα (Platelet Derived Growth Factor Receptor α), PRO (Probenecid), PVDF (polyvinylidene difluoride), S.E.M. (Standard Error of the Mean), TMZ (Temozolomide), WB (Western Blot)
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