Department of Neurosurgery, Greifswald University Medical Center, Greifswald, Germany.
ZIK plasmatis, Leibniz Institute for Plasma Science and Technology (INP), Greifswald, Germany.
Oncoimmunology. 2024 Dec 31;13(1):2425124. doi: 10.1080/2162402X.2024.2425124. Epub 2024 Nov 10.
Human glioblastoma multiforme (GBM) is a highly aggressive tumor with insufficient therapies available. Especially, novel concepts of immune therapies fail due to a complex immunosuppressive microenvironment, high mutational rates, and inter-patient variations. The intratumoral heterogeneity is currently not sufficiently investigated.
Biopsies from six different locations were taken in a cohort of 16 GBM patients who underwent surgery. The tissue slides were analyzed utilizing high-content imaging microscopy and algorithm-based image quantification. Several immune markers for macrophage and microglia subpopulations were investigated. Flow cytometry was used to validate key results. Besides the surface marker, cytokines were measured and categorized based on their heterogenicity and overall expression.
M2-like antigens, including CD204, CD163, Arg1, and CSF1R, showed comparatively higher expression, with GFAP displaying the least intratumoral heterogeneity. In contrast, anti-tumor-macrophage-like antigens, such as PSGL-1, CD16, CD68, and MHC-II, exhibited low overall expression and concurrent high intratumoral heterogeneity. CD16 and PSGL-1 were the most heterogeneous antigens. High expression levels were observed for cytokines IL-6, VEGF, and CCL-2. VILIP-a was revealed to differentiate most in principle component analysis. Cytokines with the lowest overall expression, such as TGF-β1, β-NGF, TNF-α, and TREM1, showed low intratumoral heterogeneity, in contrast to βNGF, TNF-α, and IL-18, which displayed high heterogeneity despite low expression.
The study showed high intratumoral heterogeneity in GBM, emphasizing the need for a more detailed understanding of the tumor microenvironment. The described findings could be essential for future personalized treatment strategies and the implementation of reliable diagnostics in GBM.
人胶质母细胞瘤（GBM）是一种侵袭性很强的肿瘤，目前治疗手段有限。特别是，由于免疫抑制微环境复杂、突变率高和个体间差异大，新型免疫治疗方法也未能取得成功。目前，肿瘤内异质性尚未得到充分研究。
对 16 名接受手术的 GBM 患者的六个不同部位进行活检。利用高内涵成像显微镜和基于算法的图像定量分析对组织切片进行分析。研究了几种用于巨噬细胞和小胶质细胞亚群的免疫标志物。使用流式细胞术验证了关键结果。除了表面标志物外，还根据细胞因子的异质性和整体表达进行了测量和分类。
M2 样抗原，包括 CD204、CD163、Arg1 和 CSF1R，表达较高，GFAP 的肿瘤内异质性最低。相比之下，抗肿瘤巨噬细胞样抗原，如 PSGL-1、CD16、CD68 和 MHC-II，整体表达水平较低，同时具有较高的肿瘤内异质性。CD16 和 PSGL-1 是异质性最高的抗原。IL-6、VEGF 和 CCL-2 等细胞因子表达水平较高。VILIP-a 在主成分分析中差异最大。TGF-β1、β-NGF、TNF-α 和 TREM1 等整体表达水平较低的细胞因子，肿瘤内异质性较低，而β-NGF、TNF-α 和 IL-18 尽管表达水平较低，但异质性较高。
本研究表明 GBM 存在高度的肿瘤内异质性，强调需要更详细地了解肿瘤微环境。所描述的发现对于未来的个性化治疗策略和在 GBM 中实施可靠的诊断可能至关重要。
