title: Chimeric EWSR1-FLI1 regulates the Ewing sarcoma susceptibility gene EGR2 via a GGAA microsatellite.
abstract: Deciphering the ways in which somatic mutations and germline susceptibility variants cooperate to promote cancer is challenging. Ewing sarcoma is characterized by fusions between EWSR1 and members of the ETS gene family, usually EWSR1-FLI1, leading to the generation of oncogenic transcription factors that bind DNA at GGAA motifs. A recent genome-wide association study identified susceptibility variants near EGR2. Here we found that EGR2 knockdown inhibited proliferation, clonogenicity and spheroidal growth in vitro and induced regression of Ewing sarcoma xenografts. Targeted germline deep sequencing of the EGR2 locus in affected subjects and controls identified 291 Ewing-associated SNPs. At rs79965208, the A risk allele connected adjacent GGAA repeats by converting an interspaced GGAT motif into a GGAA motif, thereby increasing the number of consecutive GGAA motifs and thus the EWSR1-FLI1-dependent enhancer activity of this sequence, with epigenetic characteristics of an active regulatory element. EWSR1-FLI1 preferentially bound to the A risk allele, which increased global and allele-specific EGR2 expression. Collectively, our findings establish cooperation between a dominant oncogene and a susceptibility variant that regulates a major driver of Ewing sarcomagenesis.
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Thomas G P
authors.initials: TG
authors.lastname: Grünewald
authors.affiliation: Institut Curie Genomics of Excellence (ICGex) Platform, Institut Curie Research Center, Paris, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Bernard
authors.affiliation: École Normale Supérieure (ENS), Institut de Biologie de l'ENS (IBENS), INSERM U1024, CNRS UMR8197, Paris, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Gilardi-Hebenstreit
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.|Institut Curie Genomics of Excellence (ICGex) Platform, Institut Curie Research Center, Paris, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Raynal
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Surdez
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Marie-Ming
authors.initials: MM
authors.lastname: Aynaud
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Mirabeau
authors.affiliation: Instituto de Investigación de Enfermedades Raras, Instituto de Salud Carlos III, Madrid, Spain.
authors.forename: Florencia
authors.initials: F
authors.lastname: Cidre-Aranaz
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Franck
authors.identifier: http://orcid.org/0000-0003-4731-7817
authors.initials: F
authors.lastname: Tirode
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Sakina
authors.initials: S
authors.lastname: Zaidi
authors.affiliation: INSERM U916 Biology of Sarcomas, Institut Bergonié, Bordeaux, France.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Perot
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Anneliene H
authors.initials: AH
authors.lastname: Jonker
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.
authors.forename: Carlo
authors.initials: C
authors.lastname: Lucchesi
authors.affiliation: Département d'Epidémiologie et de Biostatistiques, Institut Gustave Roussy, Villejuif, France.
authors.forename: Marie-Cécile
authors.initials: MC
authors.lastname: Le Deley
authors.affiliation: Département de Pédiatrie, Institut Gustave Roussy, Villejuif, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Oberlin
authors.affiliation: Institute for Pediatric Hematology and Oncology, Leon-Bérard Cancer Center, University of Lyon, Lyon, France.
authors.forename: Perrine
authors.initials: P
authors.lastname: Marec-Bérard
authors.affiliation: INSERM U1052, Léon-Bérard Cancer Centre, Cancer Research Center of Lyon, Lyon, France.
authors.forename: Amélie S
authors.initials: AS
authors.lastname: Véron
authors.affiliation: Unité Génétique Somatique (UGS), Institut Curie Centre Hospitalier, Paris, France.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Reynaud
authors.affiliation: Unité Génétique Somatique (UGS), Institut Curie Centre Hospitalier, Paris, France.
authors.forename: Eve
authors.initials: E
authors.lastname: Lapouble
authors.affiliation: INSERM U900, Bioinformatics, Biostatistics, Epidemiology and Computational Systems Biology of Cancer, Institut Curie Research Center, Paris, France.|Mines ParisTech, Fontainebleau, France.
authors.forename: Valentina
authors.initials: V
authors.lastname: Boeva
authors.affiliation: Institut Curie Genomics of Excellence (ICGex) Platform, Institut Curie Research Center, Paris, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rio Frio
authors.affiliation: Instituto de Investigación de Enfermedades Raras, Instituto de Salud Carlos III, Madrid, Spain.
authors.forename: Javier
authors.identifier: http://orcid.org/0000-0002-6287-8391
authors.initials: J
authors.lastname: Alonso
authors.affiliation: Institute for Cancer Outcomes and Survivorship, School of Medicine, University of Alabama, Birmingham, Alabama, USA.
authors.forename: Smita
authors.initials: S
authors.lastname: Bhatia
authors.affiliation: Unité Génétique Somatique (UGS), Institut Curie Centre Hospitalier, Paris, France.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Pierron
authors.affiliation: Centre de Recherche sur les Pathologies Prostatiques (CeRePP)-Laboratory for Urology, Research Team 2, UPMC, Hôpital Tenon, Paris, France.
authors.forename: Geraldine
authors.initials: G
authors.lastname: Cancel-Tassin
authors.affiliation: Centre de Recherche sur les Pathologies Prostatiques (CeRePP)-Laboratory for Urology, Research Team 2, UPMC, Hôpital Tenon, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Cussenot
authors.affiliation: INSERM U1052, Léon-Bérard Cancer Centre, Cancer Research Center of Lyon, Lyon, France.
authors.forename: David G
authors.initials: DG
authors.lastname: Cox
authors.affiliation: Division of Cancer Epidemiology and Genetics (DCEG), National Cancer Institute (NCI), Bethesda, Maryland, USA.
authors.forename: Lindsay M
authors.initials: LM
authors.lastname: Morton
authors.affiliation: Division of Cancer Epidemiology and Genetics (DCEG), National Cancer Institute (NCI), Bethesda, Maryland, USA.
authors.forename: Mitchell J
authors.initials: MJ
authors.lastname: Machiela
authors.affiliation: Division of Cancer Epidemiology and Genetics (DCEG), National Cancer Institute (NCI), Bethesda, Maryland, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Chanock
authors.affiliation: École Normale Supérieure (ENS), Institut de Biologie de l'ENS (IBENS), INSERM U1024, CNRS UMR8197, Paris, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Charnay
authors.affiliation: Genetics and Biology of Cancers Unit, Institut Curie, PSL Research University, Paris, France.|INSERM U830, Institut Curie Research Center, Paris, France.|Institut Curie Genomics of Excellence (ICGex) Platform, Institut Curie Research Center, Paris, France.|Unité Génétique Somatique (UGS), Institut Curie Centre Hospitalier, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Delattre
mesh_terms: D000818:Animals; D001483:Base Sequence; D001859:Bone Neoplasms / Q000235:genetics* / Q000473:pathology; D002338:Carotenoids / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D051767:Early Growth Response Protein 2 / Q000235:genetics*; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D006801:Humans; D016513:Mice, SCID; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D010105:Oxygenases / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics*; D040641:Quantitative Trait Loci; D034802:RNA-Binding Protein EWS / Q000235:genetics*; D012512:Sarcoma, Ewing / Q000235:genetics* / Q000473:pathology; D047368:Tumor Burden
chemical_list: C497326:EGR2 protein, human; C106031:EWS-FLI fusion protein; D051767:Early Growth Response Protein 2; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; C110027:beta-apocarotenoid-14',13'-dioxygenase; D002338:Carotenoids; D010105:Oxygenases