title: Telomerase activation by genomic rearrangements in high-risk neuroblastoma.
abstract: Neuroblastoma is a malignant paediatric tumour of the sympathetic nervous system. Roughly half of these tumours regress spontaneously or are cured by limited therapy. By contrast, high-risk neuroblastomas have an unfavourable clinical course despite intensive multimodal treatment, and their molecular basis has remained largely elusive. Here we have performed whole-genome sequencing of 56 neuroblastomas (high-risk, n = 39; low-risk, n = 17) and discovered recurrent genomic rearrangements affecting a chromosomal region at 5p15.33 proximal of the telomerase reverse transcriptase gene (TERT). These rearrangements occurred only in high-risk neuroblastomas (12/39, 31%) in a mutually exclusive fashion with MYCN amplifications and ATRX mutations, which are known genetic events in this tumour type. In an extended case series (n = 217), TERT rearrangements defined a subgroup of high-risk tumours with particularly poor outcome. Despite a large structural diversity of these rearrangements, they all induced massive transcriptional upregulation of TERT. In the remaining high-risk tumours, TERT expression was also elevated in MYCN-amplified tumours, whereas alternative lengthening of telomeres was present in neuroblastomas without TERT or MYCN alterations, suggesting that telomere lengthening represents a central mechanism defining this subtype. The 5p15.33 rearrangements juxtapose the TERT coding sequence to strong enhancer elements, resulting in massive chromatin remodelling and DNA methylation of the affected region. Supporting a functional role of TERT, neuroblastoma cell lines bearing rearrangements or amplified MYCN exhibited both upregulated TERT expression and enzymatic telomerase activity. In summary, our findings show that remodelling of the genomic context abrogates transcriptional silencing of TERT in high-risk neuroblastoma and places telomerase activation in the centre of transformation in a large fraction of these tumours.
authors.affiliation: Department of Translational Genomics, Center of Integrated Oncology Cologne-Bonn, Medical Faculty, University of Cologne, 50931 Cologne, Germany.|Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Peifer
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Falk
authors.initials: F
authors.lastname: Hertwig
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Frederik
authors.initials: F
authors.lastname: Roels
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Dreidax
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Gartlgruber
authors.affiliation: Department of Prostate Cancer Research, Institute of Pathology, Center for Integrated Oncology Cologne-Bonn, University Hospital of Bonn, 53127 Bonn, Germany.|NEO New Oncology AG, 51105 Cologne, Germany.
authors.forename: Roopika
authors.initials: R
authors.lastname: Menon
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Krämer
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh Cancer Institute (UPCI), Hillman Cancer Center, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Justin L
authors.initials: JL
authors.lastname: Roncaioli
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.
authors.forename: Frederik
authors.initials: F
authors.lastname: Sand
authors.affiliation: NEO New Oncology AG, 51105 Cologne, Germany.
authors.forename: Johannes M
authors.initials: JM
authors.lastname: Heuckmann
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.|Cologne Center for Genomics, University of Cologne, 50931 Cologne, Germany.
authors.forename: Fakhera
authors.initials: F
authors.lastname: Ikram
authors.affiliation: Institute of Biostatistics and Clinical Research, University of Münster, 48149 Münster, Germany.
authors.forename: Rene
authors.initials: R
authors.lastname: Schmidt
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Ackermann
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Engesser
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Kahlert
authors.affiliation: Department of Prostate Cancer Research, Institute of Pathology, Center for Integrated Oncology Cologne-Bonn, University Hospital of Bonn, 53127 Bonn, Germany.
authors.forename: Wenzel
authors.initials: W
authors.lastname: Vogel
authors.affiliation: Cologne Center for Genomics, University of Cologne, 50931 Cologne, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Altmüller
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Cologne Center for Genomics, University of Cologne, 50931 Cologne, Germany.|Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, 50931 Cologne, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Nürnberg
authors.affiliation: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland 20894, USA.
authors.forename: Jean
authors.initials: J
authors.lastname: Thierry-Mieg
authors.affiliation: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland 20894, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Thierry-Mieg
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.
authors.forename: Aruljothi
authors.initials: A
authors.lastname: Mariappan
authors.affiliation: NEO New Oncology AG, 51105 Cologne, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Heynck
authors.affiliation: NEO New Oncology AG, 51105 Cologne, Germany.
authors.forename: Erika
authors.initials: E
authors.lastname: Mariotti
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Kai-Oliver
authors.initials: KO
authors.lastname: Henrich
authors.affiliation: NEO New Oncology AG, 51105 Cologne, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Gloeckner
authors.affiliation: Department of Translational Genomics, Center of Integrated Oncology Cologne-Bonn, Medical Faculty, University of Cologne, 50931 Cologne, Germany.
authors.forename: Graziella
authors.initials: G
authors.lastname: Bosco
authors.affiliation: Department of Pathology, University of Kiel, 24118 Kiel, Germany.
authors.forename: Ivo
authors.initials: I
authors.lastname: Leuschner
authors.affiliation: Functional Epigenomics, University of Cologne, 50931 Cologne, Germany.
authors.forename: Michal R
authors.initials: MR
authors.lastname: Schweiger
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Larissa
authors.initials: L
authors.lastname: Savelyeva
authors.affiliation: Department of Cell Biology, Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.affiliation: Division of Theoretical Systems Biology, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Chunxuan
authors.initials: C
authors.lastname: Shao
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Emma
authors.initials: E
authors.lastname: Bell
authors.affiliation: Division of Theoretical Systems Biology, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Höfer
authors.affiliation: Computing Center, University of Cologne, 50931 Cologne, Germany.
authors.forename: Viktor
authors.initials: V
authors.lastname: Achter
authors.affiliation: Computing Center, University of Cologne, 50931 Cologne, Germany.|Department of Informatics, University of Cologne, 50931 Cologne, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Lang
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Jessica
authors.initials: J
authors.lastname: Theissen
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Volland
authors.affiliation: Division of Biostatistics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Maral
authors.initials: M
authors.lastname: Saadati
authors.affiliation: Department of Pediatric Oncology and Hematology, Charité University Medical Center Berlin, 10117 Berlin, Germany.
authors.forename: Angelika
authors.initials: A
authors.lastname: Eggert
authors.affiliation: Center for Medical Genetics, Ghent University, 9000 Ghent, Belgium.
authors.forename: Bram
authors.initials: B
authors.lastname: de Wilde
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Berthold
authors.affiliation: BGI-Shenzhen, Bei Shan Industrial Zone, Yantian District, Shenzhen, Guangdong, 518083 China.
authors.forename: Zhiyu
authors.initials: Z
authors.lastname: Peng
authors.affiliation: Center for Pharmacogenomics and Fudan-Zhangjiang Center for Clinical Genomics, State Key Laboratory of Genetic Engineering and MOE Key Laboratory of Contemporary Anthropology School of Pharmacy and School of Life Sciences, Fudan University, Shanghai 201203, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Center for Pharmacogenomics and Fudan-Zhangjiang Center for Clinical Genomics, State Key Laboratory of Genetic Engineering and MOE Key Laboratory of Contemporary Anthropology School of Pharmacy and School of Life Sciences, Fudan University, Shanghai 201203, China.
authors.forename: Leming
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Pathology, University of Cologne, 50937 Cologne, Germany.
authors.forename: Monika
authors.initials: M
authors.lastname: Ortmann
authors.affiliation: Department of Pathology, University of Cologne, 50937 Cologne, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Büttner
authors.affiliation: Department of Prostate Cancer Research, Institute of Pathology, Center for Integrated Oncology Cologne-Bonn, University Hospital of Bonn, 53127 Bonn, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Perner
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Hero
authors.affiliation: Department of Pediatric Oncology and Hematology, University Children's Hospital, 45147 Essen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Schramm
authors.affiliation: Department of Pediatric Oncology and Hematology, Charité University Medical Center Berlin, 10117 Berlin, Germany.|German Cancer Consortium (DKTK), 10117 Berlin, Germany.|German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Johannes H
authors.initials: JH
authors.lastname: Schulte
authors.affiliation: Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany.|Bioquant Center, University of Heidelberg, 69120 Heidelberg, Germany.|Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Carl
authors.initials: C
authors.lastname: Herrmann
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh Cancer Institute (UPCI), Hillman Cancer Center, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Roderick J
authors.initials: RJ
authors.lastname: O'Sullivan
authors.affiliation: Division Neuroblastoma Genomics (B087), German Cancer Research Center, 69120 Heidelberg, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Westermann
authors.affiliation: Department of Translational Genomics, Center of Integrated Oncology Cologne-Bonn, Medical Faculty, University of Cologne, 50931 Cologne, Germany.|Department of Pathology, University of Cologne, 50937 Cologne, Germany.
authors.forename: Roman K
authors.initials: RK
authors.lastname: Thomas
authors.affiliation: Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany.|Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Medical Faculty, University of Cologne, 50937 Cologne, Germany.|Max Planck Institute for Metabolism Research, 50931 Cologne, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Fischer
mesh_terms: D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D002895:Chromosomes, Human, Pair 5 / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics; D019175:DNA Methylation; D004742:Enhancer Elements, Genetic / Q000235:genetics; D004789:Enzyme Activation / Q000235:genetics; D005784:Gene Amplification / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D020868:Gene Silencing; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D007223:Infant; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000145:classification / Q000201:enzymology / Q000235:genetics* / Q000473:pathology*; D009687:Nuclear Proteins / Q000235:genetics; D015513:Oncogene Proteins / Q000235:genetics; D011379:Prognosis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D012306:Risk; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*; D014178:Translocation, Genetic / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics; D000075924:X-linked Nuclear Protein
chemical_list: D002843:Chromatin; C459801:MYCN protein, human; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D012333:RNA, Messenger; C509186:TERT protein, human; D019098:Telomerase; D004265:DNA Helicases; C092332:ATRX protein, human; D000075924:X-linked Nuclear Protein