Rapid diagnosis of IDH1-mutated gliomas by 2-HG detection with gas chromatography mass spectrometry
Rapid diagnosis of IDH1-mutated gliomas by 2-HG detection with gas chromatography mass spectrometry
复制标题
通过气相色谱质谱法检测 2-HG 快速诊断 IDH1 突变神经胶质瘤
DOI：
10.1038/s41374-018-0163-z
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发表时间：
2019-04-01
影响因子：
5
通讯作者：
Mao, Ying
中科院分区：
文献类型：
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作者：
Xu, Hao;Xia, Yu-Kun;Mao, Ying
编码异柠檬酸脱氢酶（IDH 1，2）的代谢基因在神经胶质瘤中经常发生突变。IDH的突变定义了胶质瘤的不同亚型并预测治疗反应。IDH突变具有将α-酮戊二酸（α-KG）转化为2-羟基戊二酸（2-HG）的显著的新变体活性，其现在通常被称为肿瘤代谢物和胶质瘤的生物标志物。应用PCR测序（n = 220）、免疫组化染色（n = 220）和气相色谱-质谱（GC-MS，n = 87）检测胶质瘤中IDH突变，并比较这些方法的敏感性和特异性。PCR测序和IHC染色是回顾性评估胶质瘤IDH 1突变的可靠方法，但这两种方法通常需要1-2天，这阻碍了它们在快速诊断中的应用。基于GC-MS的方法可以定性和定量检测2-HG，提供胶质瘤中IDH 1突变状态的信息，灵敏度和特异性均为100%。进一步优化的GC-MS为基础的方法（所谓的微柱法），使我们能够在40分钟内测定胶质瘤样品中的2-HG没有复杂或耗时的准备。最重要的是，2-HG/谷氨酸的比率被证明是用于确定突变状态的可靠参数。微柱法能够快速鉴定2-HG，为将来IDH 1突变胶质瘤的术中诊断提供了有前途的策略。
The metabolic genes encoding isocitrate dehydrogenase (IDH1, 2) are frequently mutated in gliomas. Mutation of IDH defines a distinct subtype of glioma and predicts therapeutic response. IDH mutation has a remarkable neomorphic activity of converting alpha-ketoglutarate (alpha-KG) to 2-hydroxyglutarate (2-HG), which is now commonly referred to as an oncometabolite and biomarker for gliomas. PCR-sequencing (n = 220), immunohistochemistry staining (IHC, n = 220), and gas chromatography mass spectrometry (GC-MS, n = 87) were applied to identify IDH mutation in gliomas, and the sensitivity and specificity of these strategies were compared. PCR-sequencing and IHC staining are reliable for retrospective assessment of IDH1 mutation in gliomas, but both methods usually take 1-2 days, which hinders their application for rapid diagnosis. GC-MS-based methods can detect 2-HG qualitatively and quantitatively, offering information on the IDH1 mutation status in gliomas with the sensitivity and specificity being 100%. Further optimization of the GC-MS based methodology (so called as the mini-column method) enabled us to determine 2-HG within 40 min in glioma samples without complex or time-consuming preparation. Most importantly, the ratio of 2-HG/glutamic acid was shown to be a reliable parameter for determination of mutation status. The mini-column method enables rapid identification of 2-HG, providing a promising strategy for intraoperative diagnosis of IDH1 -mutated gliomas in the future.