Background/Objectives: High-grade gliomas (HGGs), including glioblastomas, are among the most aggressive brain tumors due to their high intratumoral heterogeneity and extensive infiltration. Glioma stem-like cells (GSCs) frequently invade along white matter tracts such as the corpus callosum, but the molecular programs driving this region-specific invasion remain poorly defined. The aim of this study was to identify transcriptional signatures associated with GSC infiltration into the corpus callosum.Methods: We established an orthotopic xenograft model by implanting fluorescently labeled human GSCs into nude mouse brains. Tumor growth and invasion patterns were assessed using tissue clearing, light-sheet fluorescence microscopy, and histological analyses. To characterize region-specific molecular profiles, we performed microfluidic-based single-cell RNA expression analysis of 48 invasion- and stemness-related genes in cells isolated from the tumor bulk (TB) and corpus callosum (CC).Results: By six weeks post-implantation, GSCs displayed marked tropism for the corpus callosum, with distinct infiltration patterns captured by three-dimensional imaging. Single-cell gene expression profiling revealed significant differences in 7 of the 48 genes (14.6%) between TB- and CC-derived GSCs. These genes—NES,CCND1,GUSB,NOTCH1,E2F1,EGFR, andTGFB1—collectively defined a “corpus callosum invasion signature” (CC-Iv). CC-derived cells showed a unimodal, high-expression profile of CC-Iv genes, whereas TB cells exhibited bimodal distributions, suggesting heterogeneous transcriptional states. Importantly, higher CC-Iv expression correlated with worse survival in patients with low-grade gliomas.Conclusions: This multimodal approach identified a corpus callosum-specific invasion signature in glioma stem-like cells, revealing how local microenvironmental cues shape transcriptional reprogramming during infiltration. These findings provide new insights into the spatial heterogeneity of gliomas and highlight potential molecular targets for therapies designed to limit tumor spread through white matter tracts.
**背景/目的：** 高级别胶质瘤（包括胶质母细胞瘤）因其高度的瘤内异质性和广泛的侵袭性，属于最具侵袭性的脑肿瘤之一。胶质瘤干细胞样细胞常沿胼胝体等白质束侵袭，但驱动这种区域特异性侵袭的分子机制仍不明确。本研究旨在鉴定与GSC向胼胝体侵袭相关的转录特征。 **方法：** 我们通过将荧光标记的人源GSCs植入裸鼠脑内，建立了原位异种移植模型。利用组织透明化、光片荧光显微镜和组织学分析评估肿瘤生长和侵袭模式。为了表征区域特异性分子谱，我们对从肿瘤主体和胼胝体分离的细胞，进行了基于微流控技术的48个侵袭和干性相关基因的单细胞RNA表达分析。 **结果：** 植入六周后，GSCs显示出对胼胝体的显著趋向性，三维成像捕捉到其独特的侵袭模式。单细胞基因表达谱分析显示，TB来源和CC来源的GSCs在48个基因中有7个（14.6%）存在显著差异。这些基因——NES、CCND1、GUSB、NOTCH1、E2F1、EGFR和TGFB1——共同定义了一个“胼胝体侵袭特征”。CC来源的细胞显示出CC-Iv基因的单峰高表达谱，而TB细胞则呈现双峰分布，提示存在异质性的转录状态。重要的是，在低级别胶质瘤患者中，较高的CC-Iv表达与较差的生存期相关。 **结论：** 这种多模态方法鉴定了胶质瘤干细胞样细胞中胼胝体特异性的侵袭特征，揭示了局部微环境线索如何在侵袭过程中塑造转录重编程。这些发现为胶质瘤的空间异质性提供了新的见解，并突出了旨在限制肿瘤通过白质束扩散的疗法的潜在分子靶点。
肿瘤(癌症)患者之家