Abstract

We hypothesized that in glioblastoma recurring after radiotherapy, a condition whereby the brain endothelium undergoes radiation-induced senescence, tumor cells with endothelial phenotype may be relevant for tumor neovascularization. Matched glioblastoma samples obtained at primary surgery and at surgery for tumor recurrence after radiotherapy, all expressing epidermal growth factor receptor variant III (EGFRvIII), were assessed by a technique that combines fluorescent in situ hybridization (FISH) for the EGFR/CEP7 chromosomal probe with immunostaining for endothelial cells (CD31) and activated pericytes (α Smooth Muscle Actin). Five EGFRvIII-expressing paired primary/recurrent glioblastoma samples, in which the tumor cells showed EGFR/CEP7 amplification, were then assessed by CD31 and α Smooth Muscle Actin immunofluorescence. In glomeruloid bodies, the ratio between CD31+ cells with amplified EGFR/CEP7 signal and the total CD31+ cells was 0.23 ± 0.09 (mean ± sem) and 0.63 ± 0.07 in primary tumors and in recurrent ones, respectively (p < 0.002, Student-t test). In capillaries, the ratio of CD31+ cells with amplified EGFR/CEP7 over the total CD31+ cells lining the capillary lumen was 0.21 ± 0.06 (mean ± sem) and 0.42 ± 0.07 at primary surgery and at recurrence, respectively (p < 0.005, Student-t test). Expression of α Smooth Muscle Actin by cells with EGFR/CEP7 amplification was not observed. Then, in glioblastoma recurring after radiotherapy, where the brain endothelium suffers from radiation-induced cell senescence, tumor-derived endothelium plays a role in neo-vascularization.

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Acknowledgements

This study was partly funded by Associazione Italiana per la Ricerca sul Cancro (IG 2013 N.14574 to RP and IG 2014 N.15584 to LR-V).

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Author notes

  1. These authors contributed equally: Ivana De Pascalis, Liliana Morgante and Simone Pacioni.

  2. These authors jointly supervised this work: Luigi M. Larocca, Roberto Pallini.

Affiliations

  1. Institute of Neurosurgery, Università Cattolica del Sacro Cuore, Rome, Italy

    • Ivana De Pascalis
    • , Simone Pacioni
    • , Quintino Giorgio D’Alessandris
    •  & Roberto Pallini
  2. Institute of Human Anatomy, Università Cattolica del Sacro Cuore, Rome, Italy

    • Liliana Morgante
    •  & Stefano Giannetti
  3. Institute of Pathology, Università Cattolica del Sacro Cuore, Rome, Italy

    • Maurizio Martini
    •  & Luigi M. Larocca
  4. Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy

    • Lucia Ricci-Vitiani
  5. New Strategies to Inhibit Tumor Angiogenesis Program Fondazione Italiana per la Ricerca sul Cancro, Institute of Molecular Oncology Fondazione, Milan, Italy

    • Matteo Malinverno
    •  & Elisabetta Dejana

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The authors declare that they have no conflict of interest.

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Correspondence to Roberto Pallini.

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https://doi.org/10.1038/s41379-018-0046-2