Hyperactive Akt1 Signaling Increases Tumor Progression and DNA Repair in Embryonal Rhabdomyosarcoma RD Line and Confers Susceptibility to Glycolysis and Mevalonate Pathway Inhibitors
Silvia Codenotti,
Daniela Zizioli,
Luca Mignani,
Sara Rezzola,
Giovanna Tabellini,
Silvia Parolini,
Arianna Giacomini,
Michela Asperti,
Maura Poli,
Delia Mandracchia,
Marika Vezzoli,
Simona Bernardi,
Domenico Russo,
Stefania Mitola,
Eugenio Monti,
Luca Triggiani,
Davide Tomasini,
Stefano Gastaldello,
Matteo Cassandri,
Rossella Rota,
Francesco Marampon,
Alessandro Fanzani
Affiliations
Silvia Codenotti
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Daniela Zizioli
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Luca Mignani
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Sara Rezzola
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Giovanna Tabellini
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Silvia Parolini
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Arianna Giacomini
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Michela Asperti
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Maura Poli
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Delia Mandracchia
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Marika Vezzoli
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Simona Bernardi
Department of Clinical and Experimental Sciences, ASST Spedali Civili di Brescia, University of Brescia, 25123 Brescia, Italy
Domenico Russo
Department of Clinical and Experimental Sciences, ASST Spedali Civili di Brescia, University of Brescia, 25123 Brescia, Italy
Stefania Mitola
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Eugenio Monti
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
Luca Triggiani
Radiation Oncology Department, ASST Spedali Civili di Brescia, University of Brescia, 25123 Brescia, Italy
Davide Tomasini
Radiation Oncology Department, ASST Spedali Civili di Brescia, University of Brescia, 25123 Brescia, Italy
Stefano Gastaldello
Department of Physiology and Pharmacology, Karolinska Institutet, 17177 Stockholm, Sweden
Matteo Cassandri
Department of Hematology and Oncology, Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCS, 00165 Rome, Italy
Rossella Rota
Department of Hematology and Oncology, Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCS, 00165 Rome, Italy
Francesco Marampon
Department of Radiotherapy, Policlinico Umberto I, “Sapienza” University of Rome, 00161 Rome, Italy
Alessandro Fanzani
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
In pediatric rhabdomyosarcoma (RMS), elevated Akt signaling is associated with increased malignancy. Here, we report that expression of a constitutively active, myristoylated form of Akt1 (myrAkt1) in human RMS RD cells led to hyperactivation of the mammalian target of rapamycin (mTOR)/70-kDa ribosomal protein S6 kinase (p70S6K) pathway, resulting in the loss of both MyoD and myogenic capacity, and an increase of Ki67 expression due to high cell mitosis. MyrAkt1 signaling increased migratory and invasive cell traits, as detected by wound healing, zymography, and xenograft zebrafish assays, and promoted repair of DNA damage after radiotherapy and doxorubicin treatments, as revealed by nuclear detection of phosphorylated H2A histone family member X (γH2AX) through activation of DNA-dependent protein kinase (DNA-PK). Treatment with synthetic inhibitors of phosphatidylinositol-3-kinase (PI3K) and Akt was sufficient to completely revert the aggressive cell phenotype, while the mTOR inhibitor rapamycin failed to block cell dissemination. Furthermore, we found that pronounced Akt1 signaling increased the susceptibility to cell apoptosis after treatments with 2-deoxy-D-glucose (2-DG) and lovastatin, enzymatic inhibitors of hexokinase, and 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR), especially in combination with radiotherapy and doxorubicin. In conclusion, these data suggest that restriction of glucose metabolism and the mevalonate pathway, in combination with standard therapy, may increase therapy success in RMS tumors characterized by a dysregulated Akt signaling.
