title: Antitumor activity of sustained N-myc reduction in rhabdomyosarcomas and transcriptional block by antigene therapy.
abstract: OBJECTIVE
  Rhabdomyosarcomas are a major cause of cancer death in children, described with MYCN amplification and, in the alveolar subtype, transcription driven by the PAX3-FOXO1 fusion protein. Our aim was to determine the prevalence of N-Myc protein expression and the potential therapeutic effects of reducing expression in rhabdomyosarcomas, including use of an antigene strategy that inhibits transcription.
  
  
  METHODS
  Protein expression was assessed by immunohistochemistry. MYCN expression was reduced in representative cell lines by RNA interference and an antigene peptide nucleic acid (PNA) oligonucleotide conjugated to a nuclear localization signal peptide. Associated gene expression changes, cell viability, and apoptosis were analyzed in vitro. As a paradigm for antigene therapy, the effects of systemic treatment of mice with rhabdomyosarcoma cell line xenografts were determined.
  
  
  RESULTS
  High N-Myc levels were significantly associated with genomic amplification, presence of the PAX3/7-FOXO1 fusion genes, and proliferative capacity. Sustained reduction of N-Myc levels in all rhabdomyosarcoma cell lines that express the protein decreased cell proliferation and increased apoptosis. Positive feedback was shown to regulate PAX3-FOXO1 and N-Myc levels in the alveolar subtype that critically decrease PAX3-FOXO1 levels on reducing N-Myc. Pharmacologic systemic administration of the antigene PNA can eliminate alveolar rhabdomyosarcoma xenografts in mice, without relapse or toxicity.
  
  
  CONCLUSIONS
  N-Myc, with its restricted expression in non-fetal tissues, is a therapeutic target to treat rhabdomyosarcomas, and blocking gene transcription using antigene oligonucleotide strategies has therapeutic potential in the treatment of cancer and other diseases that has not been previously realized in vivo.
authors: Department of Pediatric Hematology, University of Bologna, Bologna, Italy. roberto.tonelli@unibo.it
authors: Roberto
authors: R
authors: Tonelli
authors: Alan
authors: A
authors: McIntyre
authors: Consuelo
authors: C
authors: Camerin
authors: Zoë S
authors: ZS
authors: Walters
authors: Korinne
authors: K
authors: Di Leo
authors: Joanna
authors: J
authors: Selfe
authors: Stefania
authors: S
authors: Purgato
authors: Edoardo
authors: E
authors: Missiaglia
authors: Andrea
authors: A
authors: Tortori
authors: Jane
authors: J
authors: Renshaw
authors: Annalisa
authors: A
authors: Astolfi
authors: Kathryn R
authors: KR
authors: Taylor
authors: Salvatore
authors: S
authors: Serravalle
authors: Ryan
authors: R
authors: Bishop
authors: Cristina
authors: C
authors: Nanni
authors: Linda J
authors: LJ
authors: Valentijn
authors: Andrea
authors: A
authors: Faccini
authors: Ivo
authors: I
authors: Leuschner
authors: Serena
authors: S
authors: Formica
authors: Jorge S
authors: JS
authors: Reis-Filho
authors: Valentina
authors: V
authors: Ambrosini
authors: Khin
authors: K
authors: Thway
authors: Monica
authors: M
authors: Franzoni
authors: Brenda
authors: B
authors: Summersgill
authors: Rosangela
authors: R
authors: Marchelli
authors: Patrizia
authors: P
authors: Hrelia
authors: Giorgio
authors: G
authors: Cantelli-Forti
authors: Stefano
authors: S
authors: Fanti
authors: Roberto
authors: R
authors: Corradini
authors: Andrea
authors: A
authors: Pession
authors: Janet
authors: J
authors: Shipley
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D045744:Cell Line, Tumor; D024202:Electrophoretic Mobility Shift Assay; D018628:Gene Dosage; D016259:Genes, myc / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D051379:Mice; D008819:Mice, Nude; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins / Q000235:genetics*; D015513:Oncogene Proteins / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis / Q000235:genetics; D051761:Paired Box Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D020135:Peptide Nucleic Acids / Q000494:pharmacology*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012208:Rhabdomyosarcoma / Q000235:genetics* / Q000628:therapy; D014158:Transcription, Genetic; D023041:Xenograft Model Antitumor Assays
chemical_list: C459801:MYCN protein, human; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D015514:Oncogene Proteins, Fusion; C471563:PAX3-FOXO1A fusion protein, human; D051761:Paired Box Transcription Factors; D020135:Peptide Nucleic Acids; D016271:Proto-Oncogene Proteins c-myc