title: Solid tumors of childhood display specific serum microRNA profiles.
abstract: BACKGROUND
  Serum biomarkers for diagnosis and risk stratification of childhood solid tumors would improve the accuracy/timeliness of diagnosis and reduce the need for invasive biopsies. We hypothesized that differential expression and/or release of microRNAs (miRNAs) by such tumors may be detected as altered serum miRNA profiles.
  
  
  METHODS
  We undertook global quantitative reverse transcription PCR (qRT-PCR) miRNA profiling (n = 741) on RNA from 53 serum samples, representing 33 diagnostic cases of common childhood cancers plus 20 controls. Technical confirmation was performed in a subset of 21 cases, plus four independent samples.
  
  
  RESULTS
  We incorporated robust quality control steps for RNA extraction, qRT-PCR efficiency and hemolysis quantification. We evaluated multiple methods to normalize global profiling data and identified the 'global mean' approach as optimal. We generated a panel of six miRNAs that were most stable in pediatric serum samples and therefore most suitable for normalization of targeted miRNA qRT-PCR data. Tumor-specific serum miRNA profiles were identified for each tumor type and selected miRNAs underwent confirmatory testing. We identified a panel of miRNAs (miR-124-3p/miR-9-3p/miR-218-5p/miR-490-5p/miR-1538) of potential importance in the clinical management of neuroblastoma, as they were consistently highly overexpressed in MYCN-amplified high-risk cases (MYCN-NB). We also derived candidate miRNA panels for noninvasive differential diagnosis of a liver mass (hepatoblastoma vs. combined MYCN-NB/NB), an abdominal mass (Wilms tumor vs. combined MYCN-NB/NB), and sarcoma subtypes.
  
  
  CONCLUSIONS
  This study describes a pipeline for robust diagnostic serum miRNA profiling in childhood solid tumors, and has identified candidate miRNA profiles for prospective testing.
  
  
  CONCLUSIONS
  We propose a new noninvasive method with the potential to diagnose childhood solid tumors.
authors.affiliation: Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom. Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom. University of Cambridge Department of Paediatrics, Addenbrooke's Hospital, Cambridge, United Kingdom. mjm16@cam.ac.uk nc109@cam.ac.uk.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Murray
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom.
authors.forename: Katie L
authors.initials: KL
authors.lastname: Raby
authors.affiliation: European Molecular Biology Laboratory European Bioinformatics Institute (EMBL-EBI), Hinxton, Cambridge, United Kingdom.
authors.forename: Harpreet K
authors.initials: HK
authors.lastname: Saini
authors.affiliation: Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom. Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom.
authors.forename: Shivani
authors.initials: S
authors.lastname: Bailey
authors.affiliation: Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom.
authors.forename: Sophie V
authors.initials: SV
authors.lastname: Wool
authors.affiliation: Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom.
authors.forename: Jane M
authors.initials: JM
authors.lastname: Tunnacliffe
authors.affiliation: European Molecular Biology Laboratory European Bioinformatics Institute (EMBL-EBI), Hinxton, Cambridge, United Kingdom.
authors.forename: Anton J
authors.initials: AJ
authors.lastname: Enright
authors.affiliation: Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom.
authors.forename: James C
authors.initials: JC
authors.lastname: Nicholson
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom. Department of Histopathology, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom. mjm16@cam.ac.uk nc109@cam.ac.uk.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Coleman
mesh_terms: D000293:Adolescent; D014408:Biomarkers, Tumor / Q000097:blood*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D035683:MicroRNAs / Q000097:blood*; D009369:Neoplasms / Q000097:blood*; D012334:RNA, Neoplasm / Q000097:blood*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs; D012334:RNA, Neoplasm