title: Genetic and functional diversity of propagating cells in glioblastoma.
abstract: Glioblastoma (GBM) is a lethal malignancy whose clinical intransigence has been linked to extensive intraclonal genetic and phenotypic diversity and the common emergence of therapeutic resistance. This interpretation embodies the implicit assumption that cancer stem cells or tumor-propagating cells are themselves genetically and functionally diverse. To test this, we screened primary GBM tumors by SNP array to identify copy number alterations (a minimum of three) that could be visualized in single cells by multicolor fluorescence in situ hybridization. Interrogation of neurosphere-derived cells (from four patients) and cells derived from secondary transplants of these same cells in NOD-SCID mice allowed us to infer the clonal and phylogenetic architectures. Whole-exome sequencing and single-cell genetic analysis in one case revealed a more complex clonal structure. This proof-of-principle experiment revealed that subclones in each GBM had variable regenerative or stem cell activity, and highlighted genetic alterations associated with more competitive propagating activity in vivo.
authors.affiliation: John van Geest Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0PY, UK.
authors.forename: Sara G M
authors.initials: SGM
authors.lastname: Piccirillo
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
authors.forename: Sue
authors.initials: S
authors.lastname: Colman
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
authors.forename: Nicola E
authors.initials: NE
authors.lastname: Potter
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
authors.forename: Frederik W
authors.initials: FW
authors.lastname: van Delft
authors.affiliation: Molecular Diagnostics, The Centre for Molecular Pathology, The Royal Marsden NHS Foundation Trust, London SM2 5PT, UK.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Lillis
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
authors.forename: Maria-Jose
authors.initials: MJ
authors.lastname: Carnicer
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
authors.forename: Lyndal
authors.initials: L
authors.lastname: Kearney
authors.affiliation: John van Geest Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0PY, UK; Division of Neurosurgery, Department of Clinical Neurosciences, University of Cambridge, Box 167 Addenbrooke's Hospital, Hills Road, Cambridge CB2 0QQ, UK. Electronic address: cw209@cam.ac.uk.
authors.forename: Colin
authors.initials: C
authors.lastname: Watts
authors.affiliation: Centre for Evolution and Cancer, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK. Electronic address: mel.greaves@icr.ac.uk.
authors.forename: Mel
authors.initials: M
authors.lastname: Greaves
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D056915:DNA Copy Number Variations; D018450:Disease Progression; D014644:Genetic Variation; D055106:Genome-Wide Association Study; D023281:Genomics; D005909:Glioblastoma / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D064593:Heterografts; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D059010:Single-Cell Analysis