IDH突变神经瘤中恶性转化的患者衍生细胞模型
A patient-derived cell model for malignant transformation in IDH-mutant glioma
影响因子:5.70000
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
摘要
在IDH突变的神经胶质瘤中通常可以看到恶性转化（MT）。在转型过程的早期阶段，在MT之前揭示其潜在机制和干预的研究兴趣越来越大。在这里，我们建立了一对匹配的3D细胞模型：403L，源自低度胶质瘤（LGG）和403H，通过使用IDH-突变的星形胶质细胞瘤样品衍生自High Glade胶质瘤（HGG），当时在Tumor诊断为WHO 2级（TUMOR SUFTACTION BURDANICAL AS级）（TMB）（TMB）和3.96 66 66的TMB（TMB）时，该患者是同一患者的样本。分别为70.07/mb）。两种细胞模型均验证为患者的样品保留IDH1 R132H的内源性表达。父母肿瘤的DNA甲基化谱是指LGG和HGG IDH突变胶质瘤簇。 Sox2，Nestin，GFAP，Olig2和Beta 3-微管蛋白的免疫阳性表明这两种模型的多节势。 403H更迅速地细胞浸润并在体内发展出渗透性HGG。从RNA测序分析中差异表达的基因（DEG）揭示了在403H模型中仅上调的肿瘤侵袭和相关基因。途径分析分别在403h和403L中分别显示了与上皮 - 间质转变（EMT）和Notch信号通路相关的基因的富集。基于质谱的靶向代谢组学和超极化（HP）1-13C丙酮酸内NMR分析表明，TCA循环和脂肪酸代谢有显着改变。 403h的柠檬酸盐，谷氨酰胺和2-HG水平明显更高。据我们所知，这是描述MT和Temozolomide（TMZ）诱导的IDH-突变胶质瘤中MT和Temozolomide（TMZ）诱导的高压剂表型（HMP）的一对匹配的细胞模型的开发，从而在MT/HMP期间提供了对遗传变化的遗传和代谢变化的见解。这种新型体外模型允许进一步研究MT在细胞水平上的机制。
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.