title: MYCN amplification confers enhanced folate dependence and methotrexate sensitivity in neuroblastoma.
abstract: MYCN amplification occurs in 20% of neuroblastomas and is strongly related to poor clinical outcome. We have identified folate-mediated one-carbon metabolism as highly upregulated in neuroblastoma tumors with MYCN amplification and have validated this finding experimentally by showing that MYCN amplified neuroblastoma cell lines have a higher requirement for folate and are significantly more sensitive to the antifolate methotrexate than cell lines without MYCN amplification. We have demonstrated that methotrexate uptake in neuroblastoma cells is mediated principally by the reduced folate carrier (RFC; SLC19A1), that SLC19A1 and MYCN expression are highly correlated in both patient tumors and cell lines, and that SLC19A1 is a direct transcriptional target of N-Myc. Finally, we assessed the relationship between SLC19A1 expression and patient survival in two independent primary tumor cohorts and found that SLC19A1 expression was associated with increased risk of relapse or death, and that SLC19A1 expression retained prognostic significance independent of age, disease stage and MYCN amplification. This study adds upregulation of folate-mediated one-carbon metabolism to the known consequences of MYCN amplification, and suggests that this pathway might be targeted in poor outcome tumors with MYCN amplification and high SLC19A1 expression.
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Diana T
authors: DT
authors: Lau
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Claudia L
authors: CL
authors: Flemming
authors: Department of Biology, University of Bologna, Bologna, Italy.
authors: Samuele
authors: S
authors: Gherardi
authors: Department of Biology, University of Bologna, Bologna, Italy.
authors: Giovanni
authors: G
authors: Perini
authors: Children's Hospital, Department of Pediatric Oncology and Hematology, University of Cologne and Centre for Molecular Medicine Cologne, Cologne, Germany.
authors: André
authors: A
authors: Oberthuer
authors: Children's Hospital, Department of Pediatric Oncology and Hematology, University of Cologne and Centre for Molecular Medicine Cologne, Cologne, Germany.
authors: Matthias
authors: M
authors: Fischer
authors: Division of Theoretical Bioinformatics, German Cancer Research Center, Heidelberg, Germany.
authors: Dilafruz
authors: D
authors: Juraeva
authors: Division of Theoretical Bioinformatics, German Cancer Research Center, Heidelberg, Germany.
authors: Benedikt
authors: B
authors: Brors
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Chengyuan
authors: C
authors: Xue
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Murray D
authors: MD
authors: Norris
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.|Kids Cancer Centre, Sydney Children's Hospital, Randwick, NSW, Australia.
authors: Glenn M
authors: GM
authors: Marshall
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Michelle
authors: M
authors: Haber
authors: Children's Cancer Institute Australia, Lowy Cancer Research Centre, Randwick, NSW, Australia.
authors: Jamie I
authors: JI
authors: Fletcher
authors: Faculty of Medicine, School of Women's and Children's Health, University of New South Wales, Sydney, NSW, Australia.|Research Portfolio, University of Sydney, Sydney, NSW, Australia.
authors: Lesley J
authors: LJ
authors: Ashton
mesh_terms: D045744:Cell Line, Tumor; D005492:Folic Acid / Q000378:metabolism; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D005784:Gene Amplification; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology*; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000235:genetics / Q000401:mortality / Q000473:pathology*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D058978:Reduced Folate Carrier Protein / Q000378:metabolism*
chemical_list: D005493:Folic Acid Antagonists; D047908:Intracellular Signaling Peptides and Proteins; C459801:MYCN protein, human; D000071447:N-Myc Proto-Oncogene Protein; C099927:NMI protein, human; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D012333:RNA, Messenger; D058978:Reduced Folate Carrier Protein; C548699:SLC19A1 protein, human; D005492:Folic Acid; D008727:Methotrexate
keywords: MYC; MYCN; SLC19A1; methotrexate; neuroblastoma