恶性转化 (MT) 常见于 IDH 突变神经胶质瘤。人们越来越有兴趣揭示其潜在机制并在 MT 之前在转化过程的早期阶段进行干预。在这里，我们利用来自同一患者的 IDH 突变星形细胞瘤样本建立了一对独特的匹配 3D 细胞模型：403L（源自低级别神经胶质瘤 (LGG)）和 403H（源自高级别神经胶质瘤 (HGG)）当肿瘤被诊断为WHO 2级（肿瘤突变负荷（TMB）为3.96/Mb）和后来的4级（TMB为70.07/Mb）时，分别。两种细胞模型都经过患者样本的验证，保留了 IDH1 R132H 的内源表达。亲本肿瘤的 DNA 甲基化谱称为 LGG 和 HGG IDH 突变神经胶质瘤簇。 SOX2、NESTIN、GFAP、OLIG2 和 β 3-Tubulin 的免疫阳性表明这两种模型都具有多谱系潜力。 403H更促进细胞侵袭并在体内形成浸润性HGG。 RNA测序分析的差异表达基因（DEG）揭示了403H模型中肿瘤侵袭和侵袭性相关基因专门上调。通路分析显示，403H 和 403L 中分别富集了与上皮间质转化 (EMT) 和 Notch 信号通路相关的基因。基于质谱的靶向代谢组学和超极化 (HP) 1-13C 丙酮酸细胞内 NMR 分析表明 TCA 循环和脂肪酸代谢发生了显着变化。 403H 中柠檬酸盐、谷氨酰胺和 2-HG 水平显着升高。据我们所知，这是第一份描述 IDH 突变神经胶质瘤中代表 MT 和替莫唑胺 (TMZ) 诱导的超突变表型 (HMP) 的配对 3D 患者来源细胞模型的开发的报告，提供了对遗传和代谢的见解MT/HMP 期间发生变化。这种新颖的体外模型可以在细胞水平上进一步研究 MT 的机制。© 2024。这是美国政府的作品，在美国不受版权保护；可能适用外国版权保护。

Malignant transformation (MT) is commonly seen in IDH-mutant gliomas. There has been a growing research interest in revealing its underlying mechanisms and intervening prior to MT at the early stages of the transforming process. Here we established a unique pair of matched 3D cell models: 403L, derived from a low-grade glioma (LGG), and 403H, derived from a high-grade glioma (HGG), by utilizing IDH-mutant astrocytoma samples from the same patient when the tumor was diagnosed as WHO grade 2 (tumor mutational burden (TMB) of 3.96/Mb) and later as grade 4 (TMB of 70.07/Mb), respectively. Both cell models were authenticated to a patient's sample retaining endogenous expression of IDH1 R132H. DNA methylation profiles of the parental tumors referred to LGG and HGG IDH-mutant glioma clusters. The immunopositivity of SOX2, NESTIN, GFAP, OLIG2, and beta 3-Tubulin suggested the multilineage potential of both models. 403H was more prompt to cell invasion and developed infiltrative HGG in vivo. The differentially expressed genes (DEGs) from the RNA sequencing analysis revealed the tumor invasion and aggressiveness related genes exclusively upregulated in the 403H model. Pathway analysis showcased an enrichment of genes associated with epithelial-mesenchymal transition (EMT) and Notch signaling pathways in 403H and 403L, respectively. Mass spectrometry-based targeted metabolomics and hyperpolarized (HP) 1-13C pyruvate in-cell NMR analyses demonstrated significant alterations in the TCA cycle and fatty acid metabolism. Citrate, glutamine, and 2-HG levels were significantly higher in 403H. To our knowledge, this is the first report describing the development of a matched pair of 3D patient-derived cell models representative of MT and temozolomide (TMZ)-induced hypermutator phenotype (HMP) in IDH-mutant glioma, providing insights into genetic and metabolic changes during MT/HMP. This novel in vitro model allows further investigation of the mechanisms of MT at the cellular level.© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.