Longitudinal analysis of diffuse glioma reveals cell state dynamics at recurrence associated with changes in genetics and the microenvironment
Longitudinal analysis of diffuse glioma reveals cell state dynamics at recurrence associated with changes in genetics and the microenvironment
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弥漫性胶质瘤的纵向分析揭示了与遗传学和微环境变化相关的复发时的细胞状态动态
DOI：
10.1101/2021.05.03.442486
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Varn F
中科院分区：
文献类型：
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作者：
Varn F
迄今为止，我们对胶质母细胞瘤（GBM）瘤内异质性的理解，特别是在神经发育的背景下，主要集中在更容易通过手术接近的肿瘤核心生态位。相比之下，浸润边缘的 GBM 细胞逃避手术切除并导致肿瘤复发的生物学特性仍然知之甚少。为此，我们对取自六个 GBM 肿瘤的肿瘤核心和浸润边缘的约 62,000 个细胞核进行显微解剖和单核 RNA 测序 (snRNA-seq)，这些细胞核具有不同的基因组驱动因素，包括 IDH1、EGFR、PDGFRA、FGFR3 和 NF1。无偏聚类揭示了不同的肿瘤和非肿瘤群体，并使用拷贝数变异分析进一步区分。在预测从人类成人新皮质/室下区和产前生发基质的 snRNA-seq 分析中获得的先前定义的特征后，我们发现大约 90% 的肿瘤细胞重现了神经发育样分子表型，重复了产前星形胶质细胞和星形胶质细胞和少突胶质细胞谱系分化之前的独特神经胶质中间祖细胞群 (g-IPC) 的基因表达特征。通过最可信的显微解剖（n = 4）检查样本的浸润边缘，我们发现，虽然该生态位中不同的肿瘤细胞群表达原神经和经典特征，但这些细胞相对于肿瘤核心总体富集 g-IPC 样表型，而与肿瘤的基因组改变无关。特别是位于浸润边缘的细胞子集，再现了未明确的 g-IPC 亚型的特征，表达星形胶质细胞和少突胶质细胞标记。轨迹分析还揭示了核心肿瘤细胞和边缘肿瘤细胞的不同分支，它们分别主要是星形胶质细胞样和g-IPC样细胞。浸润边缘与肿瘤核心的 GBM 细胞的差异基因表达分析揭示了以 EGFR、ERBB4、PCDH9 和 PCDH15 为主的迁移特征。最终，这种对浸润边缘异质性的高分辨率分析使我们能够发现 GBM 侵袭的潜在目标驱动因素。
Our understanding of glioblastoma (GBM) intratumoral heterogeneity, particularly in the context of neurodevelopment, has thus far been primarily focused on the more surgically accessible tumor core niche. In contrast, the biology of GBM cells at the infiltrative edge, which evade surgical resection and drive tumor recurrence, remains poorly characterized. To this end, we microdissected and performed single-nuclei RNA sequencing (snRNA-seq) on approximately 62,000 nuclei taken from the tumor core and from the infiltrative edge of six GBM tumors with diverse genomic drivers, includingIDH1,EGFR,PDGFRA,FGFR3, andNF1. Unbiased clustering reveals distinct neoplastic and non-neoplastic populations, further distinguished using copy number variation analysis. After projecting previously defined signatures taken from snRNA-seq analysis of human adult neocortex/subventricular zone and prenatal germinal matrix, we find that approximately 90% of tumor cells recapitulate a neurodevelopment-like molecular phenotype, reprising gene expression signatures of prenatal astrocytes and of a distinct glial intermediate progenitor cell population (g-IPC) that precedes both astrocyte and oligodendrocyte lineage differentiation. Examining the infiltrative edge of samples with the most confident microdissection (n=4), we see that while distinct populations of tumor cells in this niche express proneural and classical signatures, these cells are overall enriched for a g-IPC-like phenotype, relative to the tumor core, irrespective of the tumors’ genomic alterations. A subset of cells at the infiltrative edge, in particular, recapitulates the signature of an uncommitted g-IPC subtype, expressing both astroglial and oligodendroglial markers. Trajectory analyses also reveal distinct branches of core and edge tumor cells, which are predominantly astrocyte- and g-IPC-like, respectively. Differential gene expression analysis of GBM cells at the infiltrative edge vs. tumor core reveals a migration signature, dominated byEGFR, ERBB4, PCDH9,andPCDH15. Ultimately, this high resolution analysis of heterogeneity at the infiltrative edge allows us to uncover potentially targetable drivers of invasion in GBM.