Fan Kaiyu, Wei Yifan, Ou Yunwei, Gong Jian
Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100070, China.
China National Clinical Research Center for Neurological Diseases, Beijing, 100070, China.
Brain Tumor Pathol. 2023 Oct;40(4):191-203. doi: 10.1007/s10014-023-00467-4. Epub 2023 Aug 9.
To explore the characteristics of the immune microenvironment (IME) of medulloblastoma (MB) by four methods: flow cytometry (FCM), immunohistochemical (IHC), bulk RNA expression and single cell RNA sequencing (scRNA-seq), we collected the intraoperative specimens of MB, ependymoma (EPN), high-grade glioma (HGG), and low-grade glioma (LGG) to make a cross-cancer comparison. The specimens were subjected to FCM and IHC respectively, and deconvolution from bulk RNA expression data and scRNA-seq analysis were performed in MB from the GEO database. FCM and IHC analysis found that the proportion of lymphocytes (LC) and T cells between MB and other brain tumors were significantly different. The deconvolution of bulk RNA expression data showed that only the proportion of cell types in MCPCOUNTER changed greatly. scRNA-seq found that the proportion of various immune cells in the IME of MB differed between different subtypes. Techniques such as FCM, IHC, bulk RNA expression, and scRNA-seq can sort out different immune cell subsets to a certain extent and quantify their proportions. The four methods have their own strengthens and limitations, but for highly heterogeneous tumor such as MB, integrated analysis of multiple methods is a better choice.
为了通过流式细胞术（FCM）、免疫组织化学（IHC）、批量RNA表达和单细胞RNA测序（scRNA-seq）这四种方法探索髓母细胞瘤（MB）免疫微环境（IME）的特征，我们收集了MB、室管膜瘤（EPN）、高级别胶质瘤（HGG）和低级别胶质瘤（LGG）的术中标本进行跨癌比较。标本分别进行FCM和IHC检测，并对来自GEO数据库的MB进行批量RNA表达数据反卷积和scRNA-seq分析。FCM和IHC分析发现，MB与其他脑肿瘤之间淋巴细胞（LC）和T细胞的比例存在显著差异。批量RNA表达数据反卷积显示，只有MCPCOUNTER中的细胞类型比例变化很大。scRNA-seq发现，MB的IME中各种免疫细胞的比例在不同亚型之间存在差异。FCM、IHC、批量RNA表达和scRNA-seq等技术可以在一定程度上区分不同的免疫细胞亚群并量化其比例。这四种方法各有优缺点，但对于MB这种高度异质性的肿瘤，多种方法的综合分析是更好的选择。
