Single-cell profiling of D-2-hydroxyglutarate using surface-immobilized resazurin analogs.
Single-cell profiling of D-2-hydroxyglutarate using surface-immobilized resazurin analogs.
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DOI：
10.1016/j.bios.2021.113368
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发表时间：
2021-10-15
影响因子：
12.6
通讯作者：
Xue M
中科院分区：
文献类型：
--
作者：
Cheng H;Li Z;Guo Z;Shao S;Mo L;Wei W;Xue M
由于异柠檬酸脱氢酶 (IDH) 突变，D-2-羟基戊二酸 (D2HG) 作为致癌代谢物过量产生。 D2HG 的积累会导致许多酶的功能障碍和全基因组表观遗传改变，从而促进肿瘤发生。单细胞分辨率下的 D2HG 定量有助于了解 IDH 突变癌症的表型特征并确定有效的治疗方法。在这项研究中，我们开发了一种分析方法，通过在基于刃天青的荧光报告基因上采用级联酶反应来检测单个癌细胞中的 D2HG 水平。刃天青探针通过生物素-链霉亲和素相互作用固定到传感表面。这种表面化学经过合理优化，可有效地将 D2HG 水平转化为灵敏的荧光读数。该D2HG检测对D2HG表现出良好的选择性和高灵敏度，并且与之前开发的单细胞条形码芯片（SCBC）技术兼容。使用 SCBC 平台，我们对单个 IDH 突变神经胶质瘤细胞中的 D2HG、葡萄糖摄取和关键致癌信号蛋白进行同步单细胞分析。结果揭示了代谢信号和致癌信号之间复杂的相互作用，并确定了针对这些 IDH 突变神经胶质瘤细胞的有效联合靶向治疗。
D-2-hydroxyglutarate (D2HG) is over-produced as an oncometabolite due to mutations in isocitrate dehydrogenases (IDHs). Accumulation of D2HG can cause the dysfunction of many enzymes and genome-wide epigenetic alterations, which can promote oncogenesis. Quantification of D2HG at single-cell resolution can help understand the phenotypic signatures of IDH-mutant cancers and identify effective therapeutics. In this study, we developed an analytical method to detect D2HG levels in single cancer cells by adapting cascade enzymatic reactions on a resazurin-based fluorescence reporter. The resazurin probe was immobilized to the sensing surface via biotin-streptavidin interaction. This surface chemistry was rationally optimized to translate the D2HG levels to sensitive fluorescence readouts efficiently. This D2HG assay demonstrated good selectivity and high sensitivity toward D2HG, and it was compatible with the previously developed single-cell barcode chip (SCBC) technology. Using the SCBC platform, we performed simultaneous single-cell profiling of D2HG, glucose uptake, and critical oncogenic signaling proteins in single IDH-mutant glioma cells. The results unveiled the complex interplays between metabolic and oncogenic signaling and led to the identification of effective combination targeted therapy against these IDH-mutant glioma cells.
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影响因子：
29
作者：
Pavlova NN;Thompson CB
通讯作者：
Thompson CB
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
12.7
作者：
Balss, Joerg;Pusch, Stefan;von Deimling, Andreas
通讯作者：
von Deimling, Andreas
DOI：
10.1002/anie.201803034
发表时间：
2018-09-03
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Li Z;Cheng H;Shao S;Lu X;Mo L;Tsang J;Zeng P;Guo Z;Wang S;Nathanson DA;Heath JR;Wei W;Xue M
通讯作者：
Xue M
当前与IDH1突变胶质瘤背景下的癌症模型A参考的生物标志物相关程序。
影响因子：
9
作者：
Mehrjardi NZ;Hänggi D;Kahlert UD
通讯作者：
Kahlert UD