title: Hh/Gli antagonist in acute myeloid leukemia with CBFA2T3-GLIS2 fusion gene.
abstract: BACKGROUND
  CBFA2T3-GLIS2 is a fusion gene found in 17% of non-Down syndrome acute megakaryoblastic leukemia (non-DS AMKL, FAB M7) and in 8% of pediatric cytogenetically normal acute myeloid leukemia (CN-AML, in association with several French-American-British (FAB) subtypes). Children with AML harboring this aberration have a poor outcome, regardless of the FAB subtype. This fusion gene drives a peculiar expression pattern and leads to overexpression of some of Hedgehog-related genes. GLI-similar protein 2 (GLIS2) is closely related to the GLI family, the final effectors of classic Hedgehog pathway. These observations lend compelling support to the application of GLI inhibitors in the treatment of AML with the aberration CBFA2T3-GLIS2. GANT61 is, nowadays, the most potent inhibitor of GLI family proteins.
  
  
  METHODS
  We exposed to GANT61 AML cell lines and primary cells positive and negative for CBFA2T3-GLIS2 and analyzed the effect on cellular viability, induction of apoptosis, cell cycle, and expression profile.
  
  
  RESULTS
  As compared to AML cells without GLIS2 fusion, GANT61 exposure resulted in higher sensitivity of both cell lines and primary AML cells carrying CBFA2T3-GLIS2 to undergo apoptosis and G1 cell cycle arrest. Remarkably, gene expression studies demonstrated downregulation of GLIS2-specific signature genes in both treated cell lines and primary cells, in comparison with untreated cells. Moreover, chromatin immunoprecipitation analysis revealed direct regulation by GLIS2 chimeric protein of DNMT1 and DNMT3B, two genes implicated in important epigenetic functions.
  
  
  CONCLUSIONS
  Our findings indicate that the GLI inhibitor GANT61 may be used to specifically target the CBFA2T3-GLIS2 fusion gene in pediatric AML.
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Masetti
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy. salvatore.bertuccio@studio.unibo.it.
authors.forename: Salvatore Nicola
authors.identifier: http://orcid.org/0000-0002-5428-6716
authors.initials: SN
authors.lastname: Bertuccio
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.|"Giorgio Prodi" Cancer Research Center, University of Bologna, Bologna, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Astolfi
authors.affiliation: Institute of Molecular Genetics, National Research Council-IOR, Bologna, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Chiarini
authors.affiliation: Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Lonetti
authors.affiliation: "Giorgio Prodi" Cancer Research Center, University of Bologna, Bologna, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Indio
authors.affiliation: "Giorgio Prodi" Cancer Research Center, University of Bologna, Bologna, Italy.
authors.forename: Matilde
authors.initials: M
authors.lastname: De Luca
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.
authors.forename: Jessica
authors.initials: J
authors.lastname: Bandini
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Serravalle
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Franzoni
authors.affiliation: Department of Woman and Child Health, Hematology-Oncology, University of Padova, Padova, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Pigazzi
authors.affiliation: Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
authors.forename: Alberto Maria
authors.initials: AM
authors.lastname: Martelli
authors.affiliation: Department of Woman and Child Health, Hematology-Oncology, University of Padova, Padova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Basso
authors.affiliation: Department of Pediatric Hematology-Oncology, IRCCS Ospedale Bambino Gesù, Rome, Italy.|University of Pavia, Pavia, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Locatelli
authors.affiliation: Department of Pediatrics, "Lalla Seràgnoli", Hematology-Oncology Unit, Sant'Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 11, 40137, Bologna, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pession
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002648:Child; D015536:Down-Regulation / Q000187:drug effects; D053823:Hedgehog Proteins / Q000235:genetics; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000235:genetics* / Q000502:physiology; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D015514:Oncogene Proteins, Fusion / Q000187:drug effects* / Q000235:genetics; D011725:Pyridines / Q000494:pharmacology* / Q000627:therapeutic use; D011743:Pyrimidines / Q000494:pharmacology* / Q000627:therapeutic use; D012097:Repressor Proteins / Q000235:genetics*; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000235:genetics*; D000071676:Zinc Finger Protein GLI1 / Q000037:antagonists & inhibitors*
chemical_list: C115744:CBFA2T3 protein, human; C551027:GANT 61; C400551:GLI1 protein, human; C447919:GLIS2 protein, human; D053823:Hedgehog Proteins; D051741:Kruppel-Like Transcription Factors; D015514:Oncogene Proteins, Fusion; D011725:Pyridines; D011743:Pyrimidines; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; D000071676:Zinc Finger Protein GLI1
keywords: Acute megakaryoblastic leukemia; Acute myeloid leukemia; CBFA2T3-GLIS2; GANT61; Hedgehog pathway