The longitudinal transition of phenotypes is pivotal in glioblastoma treatment resistance and DNA methylation emerged as an important tool for classifying glioblastoma phenotypes. We aimed to characterize DNA methylation subclass heterogeneity during progression and assess its clinical impact. Matched tissues from 47 glioblastoma patients were subjected to DNA methylation profiling, including CpG-site alterations, tissue and serum deconvolution, mass spectrometry, and immunoassay. Effects of clinical characteristics on temporal changes and outcomes were studied. Among 47 patients, 8 (17.0%) had non-matching classifications at recurrence. In the remaining 39 cases, 28.2% showed dominant DNA methylation subclass transitions, with 72.7% being a mesenchymal subclass. In general, glioblastomas with a subclass transition showed upregulated metabolic processes. Newly diagnosed glioblastomas with mesenchymal transition displayed increased stem cell-like states and decreased immune components at diagnosis and exhibited elevated immune signatures and cytokine levels in serum. In contrast, tissue of recurrent glioblastomas with mesenchymal transition showed increased immune components but decreased stem cell-like states. Survival analyses revealed comparable outcomes for patients with and without subclass transitions. This study demonstrates a temporal heterogeneity of DNA methylation subclasses in 28.2% of glioblastomas, not impacting patient survival. Changes in cell state composition associated with subclass transition may be crucial for recurrent glioblastoma targeted therapies.

表型的纵向转变对于胶质母细胞瘤治疗耐药至关重要，DNA 甲基化成为胶质母细胞瘤表型分类的重要工具。我们的目的是表征进展过程中 DNA 甲基化亚类异质性并评估其临床影响。对 47 名胶质母细胞瘤患者的匹配组织进行了 DNA 甲基化分析，包括 CpG 位点改变、组织和血清解卷积、质谱分析和免疫分析。研究了临床特征对时间变化和结果的影响。在 47 名患者中，8 名 (17.0%) 复发时分类不匹配。在其余 39 例中，28.2% 显示显性 DNA 甲基化亚类转变，其中 72.7% 为间充质亚类。一般来说，具有亚类转变的胶质母细胞瘤表现出代谢过程上调。新诊断的具有间质转化的胶质母细胞瘤在诊断时表现出干细胞样状态增加和免疫成分减少，并且血清中免疫特征和细胞因子水平升高。相比之下，具有间质转化的复发性胶质母细胞瘤组织显示出免疫成分增加，但干细胞样状态减少。生存分析显示，有或没有亚类转变的患者的结局相当。这项研究表明 28.2% 的胶质母细胞瘤中 DNA 甲基化亚类存在时间异质性，但不会影响患者的生存。与亚类转变相关的细胞状态组成的变化可能对于复发性胶质母细胞瘤靶向治疗至关重要。