Spatial Transcriptomics of the GL261 Glioblastoma

SummaryThe GL261 syngeneic mouse model is the most widely used preclinical model for glioblastoma research, yet comprehensive spatial transcriptomic characterization of its tumor microenvironment is lacking. We performed 10x Genomics Visium spatial transcriptomics on coronal brain sections from one control and two GL261 tumor-bearing C57BL/6J mice at two weeks post-implantation. Following quality control and anatomical annotation, we identified spatial domains, performed differential expression and pathway enrichment analyses, and conducted cell type deconvolution using three independent single-cell RNA-seq references. This spatial atlas provides a framework for interpreting preclinical studies in this widely used model.     Overall designThree female C57BL/6J mice: one control (no injection, P120) and two GL261-GFP tumor-bearing (TUM1 and TUM2, implanted at P77, sacrificed at 14 and 13 days post-implantation). Fresh-frozen coronal brain sections (10 um) processed with 10x Genomics Visium Spatial Gene Expression platform. One capture area per animal.

摘要：GL261同源基因小鼠模型（GL261 syngeneic mouse model）是胶质母细胞瘤（glioblastoma）研究中应用最为广泛的临床前模型，但目前针对其肿瘤微环境的全面空间转录组学表征仍存在缺失。本研究对1只对照小鼠与2只移植后两周的GL261荷瘤C57BL/6J小鼠的冠状脑切片开展10x Genomics Visium空间转录组测序（10x Genomics Visium spatial transcriptomics）。经过质量控制与解剖注释后，我们识别了空间结构域，完成差异表达与通路富集分析，并基于三套独立的单细胞RNA测序（single-cell RNA-seq）参考数据集开展细胞类型反卷积。本空间图谱为解读这一经典临床前模型的相关临床前研究提供了研究框架。

实验整体设计：
选用3只雌性C57BL/6J小鼠：1只为对照组（未注射，P120），2只为携带GL261-GFP肿瘤的荷瘤小鼠（TUM1与TUM2，分别于P77进行肿瘤移植，分别在移植后14天、13天处死）。采用10x Genomics Visium空间基因表达平台（10x Genomics Visium Spatial Gene Expression platform）对新鲜冷冻的冠状脑切片（厚度10 μm）进行处理，每只动物设置一个捕获区域。