Chen Yicheng, Zhao Yiqing, Bajor David L, Wang Zhenghe, Selfridge J Eva
Department of Genetics and Genome Sciences, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA; Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA.
Department of Medicine, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA; Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA; Seidman Cancer Center, University Hospitals Cleveland Medical Center, 11100 Euclid Avenue, Cleveland, OH, 44106, USA.
J Genet Genomics. 2020 Jul 20;47(7):389-395. doi: 10.1016/j.jgg.2020.06.001. Epub 2020 Jun 16.
Many cancer types reprogram their metabolism to become addicted to glutamine. One of the critical enzymes in the utilization of glutamine in these cells is glutaminase. CB-839 (telaglenastat) is a drug that targets glutaminase that is currently being evaluated in many clinical trials for efficacy in various cancer types that are known to be driven by glutamine metabolism. Despite its use, there are limited assays available for testing the pharmacodynamic on-target effects of CB-839 on the limited, small-volume patient samples that are obtained in early-phase clinical trials. Thus, we developed an assay based on the cellular thermal shift assay technique using AlphaLISA technology to show that CB-839 specifically engages glutaminase in colon cancer cell lines in vitro and in minute quantities of mouse xenograft tumors. Notably, we show that this assay detects CB-839 binding to glutaminase in platelets of patients collected while receiving CB-839 on a clinical trial. This assay may be used to study the pharmacodynamic profile of CB-839 in very small tissue samples obtained from patients on a clinical trial and may be useful in future studies designed to screen other inhibitors of glutaminase.
许多癌症类型会重新编程其代谢，变得依赖谷氨酰胺。这些细胞利用谷氨酰胺的关键酶之一是谷氨酰胺酶。CB - 839（替拉格伦司他）是一种靶向谷氨酰胺酶的药物，目前正在多项临床试验中评估其对多种已知由谷氨酰胺代谢驱动的癌症类型的疗效。尽管已使用该药物，但对于在早期临床试验中获取的有限的小体积患者样本，用于测试CB - 839药效学靶点效应的检测方法却很有限。因此，我们开发了一种基于细胞热位移分析技术并使用AlphaLISA技术的检测方法，以表明CB - 839在体外和极少量小鼠异种移植肿瘤的结肠癌细胞系中特异性结合谷氨酰胺酶。值得注意的是，我们证明该检测方法能检测到在临床试验中接受CB - 839治疗的患者血小板中CB - 839与谷氨酰胺酶的结合。该检测方法可用于研究从临床试验患者获取的极少量组织样本中CB - 839的药效学特征，并且可能对未来旨在筛选其他谷氨酰胺酶抑制剂的研究有用。
