Pharmacological targeting of the ephrin receptor kinase signalling by GLPG1790 in vitro and in vivo reverts oncophenotype, induces myogenic differentiation and radiosensitizes embryonal rhabdomyosarcoma cells

Journal of Hematology & Oncology

Roberto Maggio¹  Claudio Festuccia²  Giovanni Luca Gravina²  Francesco Marampon²  Vincenzo Desiderio³  Cinzia Marchese⁴  Simona Ceccarelli⁴  Antonio Pizzuti⁴  Simona Camero⁵  Heather P. McDowell⁵  Francesca Megiorni⁵  Carlo Dominici⁵  Rajeev Shukla⁶  Francesca De Felice⁷  Vincenzo Tombolini⁷  Federica Papaccio⁸  Laurent Saniere⁹  Philippe Pujuguet⁹  Ellen Van der Aar^1,10  Filip Beirinckx^1,10  Andrea Del Fattore^1,11 

[1] Department of Biotechnological and Applied Clinical Sciences, Division of Pharmacology, University of L’Aquila;Department of Biotechnological and Applied Clinical Sciences, Division of Radiation Oncology, University of L’Aquila;Department of Experimental Medicine, Section of Biotechnology and Medical Histology and Embriology, Second University of Naples;Department of Experimental Medicine, “Sapienza” University of Rome;Department of Paediatrics and Infantile Neuropsychiatry, “Sapienza” University of Rome;Department of Perinatal and Paediatric Pathology, Alder Hey Children’s NHS Foundation Trust;Department of Radiological, Oncological and Pathological Sciences, “Sapienza” University of Rome;Division of Medical Oncology, Department of Clinical and Experimental Medicine and Surgery “F. Magrassi A. Lanzara”, Second University of Naples;Galapagos France;Galapagos NV, Industriepark Mechelen Noord;Multi-Factorial Disease and Complex Phenotype Research Area, Bambino Gesu Children’s Hospital, IRCCS;

关键词: Rhabdomyosarcoma;    GLPG1790;    Ephrin;    Cancer stem cell;    Radiation therapy;    Tumour xenografts;   

DOI  :  10.1186/s13045-017-0530-z

来源: DOAJ

【 摘 要 】

Abstract Background EPH (erythropoietin-producing hepatocellular) receptors are clinically relevant targets in several malignancies. This report describes the effects of GLPG1790, a new potent pan-EPH inhibitor, in human embryonal rhabdomyosarcoma (ERMS) cell lines. Methods EPH-A2 and Ephrin-A1 mRNA expression was quantified by real-time PCR in 14 ERMS tumour samples and in normal skeletal muscle (NSM). GLPG1790 effects were tested in RD and TE671 cell lines, two in vitro models of ERMS, by performing flow cytometry analysis, Western blotting and immunofluorescence experiments. RNA interfering experiments were performed to assess the role of specific EPH receptors. Radiations were delivered using an x-6 MV photon linear accelerator. GLPG1790 (30 mg/kg) in vivo activity alone or in combination with irradiation (2 Gy) was determined in murine xenografts. Results Our study showed, for the first time, a significant upregulation of EPH-A2 receptor and Ephrin-A1 ligand in ERMS primary biopsies in comparison to NSM. GLPG1790 in vitro induced G1-growth arrest as demonstrated by Rb, Cyclin A and Cyclin B1 decrease, as well as by p21 and p27 increment. GLPG1790 reduced migratory capacity and clonogenic potential of ERMS cells, prevented rhabdosphere formation and downregulated CD133, CXCR4 and Nanog stem cell markers. Drug treatment committed ERMS cells towards skeletal muscle differentiation by inducing a myogenic-like phenotype and increasing MYOD1, Myogenin and MyHC levels. Furthermore, GLPG1790 significantly radiosensitized ERMS cells by impairing the DNA double-strand break repair pathway. Silencing of both EPH-A2 and EPH-B2, two receptors preferentially targeted by GLPG1790, closely matched the effects of the EPH pharmacological inhibition. GLPG1790 and radiation combined treatments reduced tumour mass by 83% in mouse TE671 xenografts. Conclusions Taken together, our data suggest that altered EPH signalling plays a key role in ERMS development and that its pharmacological inhibition might represent a potential therapeutic strategy to impair stemness and to rescue myogenic program in ERMS cells.

【 授权许可】

Unknown