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.

胶质母细胞瘤（GBM）是一种致命的恶性肿瘤，其临床顽固性与广泛的克隆内遗传和表型多样性以及普遍出现的治疗耐药性有关。这种解释体现了一个隐含的假设，即癌症干细胞或肿瘤增殖细胞本身在遗传和功能上是多样化的。为了测试这一点，我们通过 SNP 阵列筛选原发性 GBM 肿瘤，以确定可以通过多色荧光原位杂交在单细胞中可视化的拷贝数改变（至少三个）。对神经球来源的细胞（来自四名患者）和 NOD-SCID 小鼠中这些相同细胞的二次移植来源的细胞的询问使我们能够推断出克隆和系统发育结构。在一个案例中，全外显子组测序和单细胞遗传分析揭示了更复杂的克隆结构。这项原理验证实验揭示了每个 GBM 中的亚克隆具有可变的再生或干细胞活性，并强调了与体内更具竞争性的繁殖活性相关的遗传改变。