Vol.:(0123456789) 1 3 Journal of Cancer Research and Clinical Oncology https://doi.org/10.1007/s00432-018-2800-8 ORIGINAL ARTICLE – CANCER RESEARCH Histone deacetylase inhibitor ITF2357 (givinostat) reverts transformed phenotype and counteracts stemness in in vitro and in vivo models of human glioblastoma Francesco Marampon 1  · Flavio Leoni 2  · Andrea Mancini 3  · Ilaria Pietrantoni 4  · Silvia Codenotti 5  · Ferella Letizia 3,6  · Francesca Megiorni 7  · Giuliana Porro 2  · Elisabetta Galbiati 2  · Pietro Pozzi 2  · Paolo Mascagni 2  · Alfredo Budillon 8  · Roberto Maggio 4  · Vincenzo Tombolini 1  · Alessandro Fanzani 5  · Giovanni Luca Gravina 3,6  · Claudio Festuccia 3 Received: 4 September 2018 / Accepted: 17 November 2018 © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract Purpose Aberrant expression and activity of histone deacetylases (HDACs) sustain glioblastoma (GBM) onset and progres- sion, and, therefore, HDAC inhibitors (HDACi) represent a promising class of anti-tumor agents. Here, we analyzed the efects of ITF2357 (givinostat), a pan-HDACi, in GBM models for its anti-neoplastic potential. Methods A set of GBM- and patient-derived GBM stem-cell lines was used and the ITF2357 efects on GBM oncophenotype were investigated in in vitro and in vivo xenograft models. Results ITF2357 inhibited HDAC activity and afected GBM cellular fate in a dose-dependent manner by inducing G 1 /S growth arrest (1–2.5 µM) or caspase-mediated cell death (≥ 2.5 µM). Chronic treatment with low doses (≤ 1 µM) induced autophagy-mediated cell death, neuronal-like phenotype, and the expression of diferentiation markers, such as glial fbrillar actin protein (GFAP) and neuron-specifc class III beta-tubulin (Tuj-1); this reduces neurosphere formation from patient- derived GBM stem cells. Autophagy inhibition counteracted the ITF2357-induced expression of diferentiation markers in p53-expressing GBM cells. Finally, in in vivo experiments, ITF2357 efciently passed the blood–brain barrier, so rapidly reaching high concentration in the brain tissues, and signifcantly afected U87MG and U251MG growth in orthotopic xenotransplanted mice. Conclusions The present fndings provide evidence of the key role played by HDACs in sustaining transformed and stem phenotype of GBM and strongly suggest that ITF2357 may have a clinical potential for the HDACi-based therapeutic strate- gies against GBM. Keywords Glioblastoma · Cancer stem cells · HDACs · HDACs’ inhibitor · Givinostat · ITF2357 Introduction Glioblastoma (GB) accounts more than 60% of all primary brain tumors. Despite multimodality therapeutic approaches, consisting of surgical debulking followed by radiotherapy combined to temozolomide chemotherapy, the median survival is 12–15 months and all new therapeutic strategies have failed to prolong survival (Johnson and O’Neill 2012). Several genetic and epigenetic alterations have been related to GBM onset, progression, and resistance to therapies, with increasing evidences focused on the role of aberrant histone acetylation in tumor etiopathogenesis (Nagarajan and Costello 2009). Histones, nuclear proteins that package and order the DNA into structural units called nucleosomes, modifying the condensed chromatin status, affect gene transcription (Chen et al. 2015). Histone acetyltransferases (HATs), by acetylating the histones at specifc aminoacid positions, open the condensed chromatin structure, thus, promoting gene transcription, whilst histone deacetylases (HDACs), by deacetylating histone tails, promote opposite efects (Chen et al. 2015). Consistent with their importance Francesco Marampon and Flavio Leoni have contributed equally. Giovanni Luca Gravina and Claudio Festuccia have contributed equally. * Claudio Festuccia claudio.festuccia@univaq.it Extended author information available on the last page of the article