title: Role of microRNAs in epigenetic silencing of the CHD5 tumor suppressor gene in neuroblastomas.
abstract: Neuroblastoma (NB), a tumor of the sympathetic nervous system, is the most common extracranial solid tumor of childhood. We and others have identified distinct patterns of genomic change that underlie diverse clinical behaviors, from spontaneous regression to relentless progression. We first identified CHD5 as a tumor suppressor gene that is frequently deleted in NBs. Mutation of the remaining CHD5 allele is rare in these tumors, yet expression is very low or absent, so expression is likely regulated by epigenetic mechanisms. In order to understand the potential role of miRNA regulation of CHD5 protein expression in NBs, we examined all miRNAs that are predicted to target the 3'-UTR using miRanda, TargetScan and other algorithms. We identified 18 miRNAs that were predicted by 2 or more programs: miR-204, -211, -216b, -17, -19ab, -20ab, -93, -106ab, -130ab, -301ab, -454, -519d, -3666. We then performed transient transfections in two NB cell lines, NLF (MYCN amplified) and SY5Y (MYCN non-amplified), with the reporter plasmid and miRNA mimic, as well as appropriate controls. We found seven miRNAs that significantly downregulated CHD5 expression in NB: miR-211, 17, -93, -20b, -106b, -204, and -3666. Interestingly, MYCN upregulates several of the candidates we identified: miR-17, -93, -106b & -20b. This suggests that miRNAs driven by MYCN and other genes represent a potential epigenetic mechanism to regulate CHD5 expression.
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Koumudi
authors.initials: K
authors.lastname: Naraparaju
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Venkatadri
authors.initials: V
authors.lastname: Kolla
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Tiangang
authors.initials: T
authors.lastname: Zhuang
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Higashi
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Iyer
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Sriharsha
authors.initials: S
authors.lastname: Kolla
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Erin R
authors.initials: ER
authors.lastname: Okawa
authors.affiliation: Department of Pediatrics, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Gerd A
authors.initials: GA
authors.lastname: Blobel
authors.affiliation: Division of Oncology and Hematology, The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Garrett M
authors.initials: GM
authors.lastname: Brodeur
mesh_terms: D045744:Cell Line, Tumor; D004265:DNA Helicases / Q000096:biosynthesis / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology*; D016147:Genes, Tumor Suppressor / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics*; D009447:Neuroblastoma / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D009419:Nerve Tissue Proteins; D004265:DNA Helicases; C471926:CHD5 protein, human
keywords: CHD5; MYCN; miRNA; neuroblastoma; tumor suppressor