IDH突变胶质瘤中恶性转化的患者源细胞模型

A patient-derived cell model for malignant transformation in IDH-mutant glioma

Acta Neuropathologica Communications

影响因子:5.7

分区:医学1区 Top / 神经科学1区

发表日期:2024 Sep 10

作者: Olga Kim, Zach Sergi, Guangyang Yu, Kazutoshi Yamamoto, Martha Quezado, Zied Abdullaev, Danel R Crooks, Shun Kishimoto, Qi Li, Peng Lu, Burchelle Blackman, Thorkell Andresson, Xiaolin Wu, Bao Tran, Jun S Wei, Wei Zhang, Meili Zhang, Hua Song, Javed Khan, Murali C Krishna, Jeffrey R Brender, Jing Wu

DOI: 10.1186/s40478-024-01860-6

摘要

恶性转化（MT）在IDH突变胶质瘤中常见。越来越多的研究关注其潜在机制及在转化早期干预。我们建立了一对独特的匹配3D细胞模型：403L，来源于低级别胶质瘤（LGG），由同一患者的肿瘤在诊断为WHO 2级（突变负荷（TMB）为3.96/Mb）时采集；以及403H，来源于高级别胶质瘤（HGG），在肿瘤诊断为WHO 4级（TMB为70.07/Mb）时采集。两者模型均确认为保持内源性IDH1 R132H表达的患者样本。亲本肿瘤的DNA甲基化谱分别对应LGG和HGG IDH突变胶质瘤簇。免疫组化检测显示SOX2、NESTIN、GFAP、OLIG2和beta 3-Tubulin呈阳性，表明两模型具有多谱系潜能。403H模型更易侵袭，并在体内形成浸润性HGG。RNA测序的差异表达基因（DEGs）显示，403H模型中特异性上调了与肿瘤侵袭和侵袭性相关的基因。通路分析显示，403H模型中富集了上皮-间充质转化（EMT）相关基因，而403L模型中富集了Notch信号通路相关基因。质谱靶向代谢组学和高极化（HP）1-13C丙酮酸细胞内NMR分析显示TCA循环和脂肪酸代谢发生显著变化。403H模型中柠檬酸、谷氨酰胺和2-HG水平显著升高。据我们所知，这是首个报道用匹配的患者来源3D细胞模型描述IDH突变胶质瘤中MT和TMZ诱导的超突变表型（HMP）的研究，为MT/HMP期间的遗传和代谢变化提供了新见解。这一新型体外模型有助于进一步研究细胞水平的MT机制。

Abstract

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.