ANGI-03. THE MIGRATORY SWITCH - INVESTIGATING MESENCHYMAL-AMOEBOID TRANSITION (MAT) IN HIGH GRADE GLIOMAS
ANGI-03. THE MIGRATORY SWITCH - INVESTIGATING MESENCHYMAL-AMOEBOID TRANSITION (MAT) IN HIGH GRADE GLIOMAS
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安吉-03。
DOI：
10.1093/neuonc/nox168.082
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发表时间：
2017
期刊：
影响因子：
15.9
通讯作者：
Taylor S
中科院分区：
文献类型：
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作者：
Taylor S
胶质母细胞瘤（GBM）是一种多克隆肿瘤，其分子异常在时间和空间上变化（肿瘤内异质性），是治疗耐药的一种机制。荧光引导的神经外科切除术是在预先给予5-氨基乙酰丙酸（5-ALA）的情况下进行的，导致单独的荧光肿瘤细胞混合在“纯”肿瘤以外的侵袭区域的非肿瘤细胞背景群体中。我们已经通过基于5-ALA的荧光激活细胞分选（FACS）分离了这个关键的侵袭性群体，以研究侵袭性GBM，而没有压倒性的背景“正常”信号。还对来自三个不同GBM患者的14个胶质瘤样品进行了全基因组基因表达分析（Affytron），通过qPCR和免疫组织化学进行验证。针对肿瘤区域和荧光分离的侵袭性细胞生成高分辨率RNAseq谱。通过线性模型的统计分析鉴定了FACS阳性细胞和其他肿瘤区域之间的325个差异表达基因（多重检验调整的P值< 0.05），其中，50个基因在所有比较中上调。这些转录组变化协调MAPK（DUSP 1，DUSP 2，DUSP 10和FOSB），趋化因子信号传导途径（CXCL 2，CXCL 3，CCL 20和NFKB 1）和细胞增殖的负调控（EREG和KLF 6）。相反，29个下调基因在FACS阳性细胞中富集信号转导（DDR2和MTSS 1 L）和ECM-受体相互作用（COL 4A 1，COL 4A 2和HSPG2.CONCLUSIONResidual GBM细胞浸润区显示出不同的分子途径和生物过程的激活。这些独特的分子特征为开发更有效的靶向治疗提供了希望，这些靶向治疗专注于这一人群，而不是大多数历史分析的主题大块肿瘤。
INTRODUCTIONGlioblastoma (GBM) is a polyclonal tumor with molecular abnormalities varying temporo-spatially (intra-tumor heterogeneity), one mechanism of therapy resistance. Fluorescence guided neurosurgical resection is performed with prior administration of 5-aminolevulinic acid (5-ALA) leading to individually fluorescent tumor cells mixed within a background population of non-neoplastic cells in the invasive region beyond “pure” tumor. We have isolated this critical invasive population by 5ALA-based fluorescence activated cell sorting (FACS) to study invasive GBM without an overwhelming background “normal” signal.METHODWe performed high-resolution RNAseq analysis on 45 specimens from 10 GBM patients including samples from tumor core, rim, invasive zone and GBM cells from within brain parenchyma by 5-ALA based FACS. Genome-wide gene expression analysis (Affymetrix) on 14 glioma samples from three different GBM patients was also undertaken, with validation by qPCR and immunohistochemistry.RESULTSFluorescent cancer cells accounted for an average of 1% of the samples subjected to FACS. High-resolution RNAseq profiles were generated for tumor regions and fluorescently isolated invasive cells. Statistical analysis by linear models identified 325 differentially expressed genes between FACS positive cells and other tumor regions (multiple test adjusted P-value < 0.05), of these, 50 genes were upregulated in all comparisons. These transcriptomic changes orchestrate MAPK (DUSP1, DUSP2, DUSP10 and FOSB), chemokine signaling pathways (CXCL2, CXCL3, CCL20 and NFKB1) and negative regulation of cell proliferation (EREG and KLF6). In contrast, 29 downregulated genes in FACS positive cells were enriched with signal transduction (DDR2 and MTSS1L) and ECM-receptor interaction (COL4A1, COL4A2, and HSPG2).CONCLUSIONResidual GBM cells in the invasive zone are shown to exhibit activation of distinct molecular pathways and biological processes. These unique molecular features offer hope for developing more efficacious targeted therapies focusing on this population rather than the bulk tumor that has been the subject of most historical analyses.