对大鼠肉瘤病毒 (Ras)-G12C 抑制剂的临床耐药性是一个挑战。癌细胞亚群已被证明会经历基因组和转录改变以促进耐药性，但在单细胞水平上响应这些药物对 Ras 信号传导的直接适应性影响尚不清楚。在这里，我们使用 Ras 生物传感器在单细胞水平上分析内源 Ras 的活性和信号环境。我们发现，用 Ras-G12C-鸟苷二磷酸抑制剂处理的 KRas-G12C 细胞亚群分别经历了由高尔基体野生型 Ras 和线粒体突变型 KRas 驱动的适应性信号传导和代谢变化。我们的 Ras 生物传感器通过 Ras 信号通路成分和代谢通道的支架，将主要穹窿蛋白识别为 Ras 激活的介质。总体而言，包括我们在内的促进单细胞水平直接分析的方法可以报告细胞亚群对癌症治疗的反应所采取的适应性，从而提供对耐药性的深入了解。© 2024。作者。

Clinical resistance to rat sarcoma virus (Ras)-G12C inhibitors is a challenge. A subpopulation of cancer cells has been shown to undergo genomic and transcriptional alterations to facilitate drug resistance but the immediate adaptive effects on Ras signaling in response to these drugs at the single-cell level is not well understood. Here, we used Ras biosensors to profile the activity and signaling environment of endogenous Ras at the single-cell level. We found that a subpopulation of KRas-G12C cells treated with Ras-G12C-guanosine-diphosphate inhibitors underwent adaptive signaling and metabolic changes driven by wild-type Ras at the Golgi and mutant KRas at the mitochondria, respectively. Our Ras biosensors identified major vault protein as a mediator of Ras activation through its scaffolding of Ras signaling pathway components and metabolite channels. Overall, methods including ours that facilitate direct analysis on the single-cell level can report the adaptations that subpopulations of cells adopt in response to cancer therapies, thus providing insight into drug resistance.© 2024. The Author(s).