Bedia Jacob S, Delgado-Gonzalez Antonio, Huang Ying-Wen, Gonzalez Veronica D, Funingana Ionut-Gabriel, Rahil Zainab, Mike Alyssa, Lowber Alexis, Vias Maria, Ashworth Alan, Brenton James D, Fantl Wendy J
Department of Urology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Department of Urology, Stanford University School of Medicine, Stanford, CA 94305, USA; Baxter Laboratory for Stem Cell Biology, Department of Microbiology & Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Cell Rep Med. 2025 Sep 16;6(9):102295. doi: 10.1016/j.xcrm.2025.102295. Epub 2025 Aug 22.
Tubo-ovarian high-grade serous carcinoma (HGSC), the most lethal gynecologic malignancy, initially responds to platinum-based chemotherapy, but due to frequent defects in the DNA damage response (DDR), most tumors develop resistance. The molecular mechanisms underlying clinical platinum resistance remain poorly defined with no biomarkers or targeted therapies to improve outcomes. Here, applying mass cytometry, we quantify phosphorylation and abundance of DDR proteins in carboplatin-treated HGSC cell line models. Despite similar levels of intranuclear platinum, a proxy for carboplatin uptake, cells follow divergent fates, reflecting DDR heterogeneity. Unsupervised analysis reveals a continuum of DDR states, and matrix factorization identifies eight protein modules. The activity of one module, containing canonical DDR proteins, increases in carboplatin-sensitive cells. Resistant cells engage a broader DDR protein module. These findings demonstrate the ability of single-cell proteomics to identify functional DDR states and reveal a DDR sensitivity module as a promising biomarker for clinical stratification and therapeutic decisions in HGSC.
输卵管卵巢高级别浆液性癌（HGSC）是最致命的妇科恶性肿瘤，最初对铂类化疗有反应，但由于DNA损伤反应（DDR）频繁出现缺陷，大多数肿瘤会产生耐药性。临床铂耐药的分子机制仍不清楚，没有生物标志物或靶向治疗方法来改善治疗结果。在此，我们应用质谱流式细胞术，对卡铂处理的HGSC细胞系模型中DDR蛋白的磷酸化和丰度进行定量。尽管细胞核内铂的水平相似（卡铂摄取的一个指标），但细胞却有不同的命运，这反映了DDR的异质性。无监督分析揭示了DDR状态的连续性，矩阵分解识别出八个蛋白质模块。一个包含经典DDR蛋白的模块的活性在对卡铂敏感的细胞中增加。耐药细胞涉及更广泛的DDR蛋白模块。这些发现证明了单细胞蛋白质组学识别功能性DDR状态的能力，并揭示了一个DDR敏感性模块作为HGSC临床分层和治疗决策的有前景的生物标志物。
