Related Experiment Video
Updated: Sep 10, 2025
Visualizing DNA Damage Repair Proteins in Patient-Derived Ovarian Cancer Organoids via Immunofluorescence Assays
Published on: February 24, 2023
Coordinated protein modules define DNA damage responses to carboplatin at single-cell resolution in human ovarian
Jacob S Bedia1, Antonio Delgado-Gonzalez2, Ying-Wen Huang1
1Department of Urology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Abstract:
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.
Insights
Tubo-ovarian high-grade serous carcinoma (HGSC) resistance to carboplatin chemotherapy is linked to DNA damage response (DDR) heterogeneity. Identifying a DDR sensitivity module could improve patient stratification and treatment for this lethal gynecologic malignancy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Tubo-ovarian high-grade serous carcinoma (HGSC) is a lethal gynecologic malignancy.
- Initial response to platinum chemotherapy is often followed by acquired resistance.
- Mechanisms of platinum resistance and effective biomarkers remain poorly defined.
Purpose of the Study:
- To investigate the heterogeneity of DNA damage response (DDR) in HGSC cell lines treated with carboplatin.
- To identify molecular mechanisms underlying carboplatin resistance in HGSC.
- To explore the potential of DDR protein modules as biomarkers for clinical stratification.
Main Methods:
- Mass cytometry was used to quantify phosphorylation and abundance of DDR proteins.
- HGSC cell line models were treated with carboplatin.
- Unsupervised analysis and matrix factorization were applied to identify DDR states and protein modules.
Main Results:
- Carboplatin-treated HGSC cells exhibited divergent fates despite similar platinum uptake, indicating DDR heterogeneity.
- A continuum of DDR states was identified, with eight distinct protein modules characterized.
- A module of canonical DDR proteins was upregulated in carboplatin-sensitive cells, while resistant cells engaged a broader DDR network.
Conclusions:
- Single-cell proteomics can effectively identify functional DDR states in HGSC.
- A specific DDR sensitivity module shows promise as a biomarker for predicting response to carboplatin.
- These findings may guide clinical stratification and therapeutic decisions for HGSC patients.
Related Concept Videos
02:37DNA Damage Can Stall the Cell Cycle
02:37DNA Damage can Stall the Cell Cycle
02:53Adaptive Mechanisms in Cancer Cells
Some of the advantages that cancer cells have on normal cells include - enhanced ability to divide without terminally differentiating, induce new blood vessel formation,...