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Phenotypic Characterization by Mass Cytometry of the Microenvironment in Ovarian Cancer and Impact of Tumor
Shamundeeswari Anandan1,2, Liv Cecilie V Thomsen1,2, Stein-Erik Gullaksen1
1Centre for Cancer Biomarkers, Department of Clinical Science, University of Bergen, 5021 Bergen, Norway.
Abstract:
Improved molecular dissection of the tumor microenvironment (TME) holds promise for treating high-grade serous ovarian cancer (HGSOC), a gynecological malignancy with high mortality. Reliable disease-related biomarkers are scarce, but single-cell mapping of the TME could identify patient-specific prognostic differences. To avoid technical variation effects, however, tissue dissociation effects on single cells must be considered. We present a novel Cytometry by Time-of-Flight antibody panel for single-cell suspensions to identify individual TME profiles of HGSOC patients and evaluate the effects of dissociation methods on results. The panel was developed utilizing cell lines, healthy donor blood, and stem cells and was applied to HGSOC tissues dissociated by six methods. Data were analyzed using Cytobank and X-shift and illustrated by t-distributed stochastic neighbor embedding plots, heatmaps, and stacked bar and error plots. The panel distinguishes the main cellular subsets and subpopulations, enabling characterization of individual TME profiles. The dissociation method affected some immune (n = 1), stromal (n = 2), and tumor (n = 3) subsets, while functional marker expressions remained comparable. In conclusion, the panel can identify subsets of the HGSOC TME and can be used for in-depth profiling. This panel represents a promising profiling tool for HGSOC when tissue handling is considered.
Insights
A new antibody panel aids in dissecting the tumor microenvironment (TME) of high-grade serous ovarian cancer (HGSOC). This tool helps identify patient-specific differences, but tissue dissociation methods can impact results.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- High-grade serous ovarian cancer (HGSOC) has high mortality, necessitating improved molecular dissection of the tumor microenvironment (TME).
- Reliable biomarkers for HGSOC are scarce, and single-cell mapping of the TME may reveal prognostic differences.
- Tissue dissociation methods can introduce technical variations affecting single-cell analysis.
Purpose of the Study:
- To develop a novel Cytometry by Time-of-Flight (CTOF) antibody panel for single-cell suspensions.
- To identify individual TME profiles in HGSOC patients.
- To evaluate the impact of different tissue dissociation methods on TME profiling.
Main Methods:
- Developed a CTOF antibody panel using cell lines, healthy donor blood, and stem cells.
- Applied the panel to HGSOC tissues dissociated by six distinct methods.
- Analyzed data using Cytobank and X-shift, visualized with t-SNE plots, heatmaps, and bar plots.
Main Results:
- The CTOF panel successfully distinguished major cellular subsets and subpopulations within the HGSOC TME.
- Dissociation methods influenced specific immune, stromal, and tumor cell subsets.
- Functional marker expression remained largely comparable across dissociation methods.
Conclusions:
- The developed CTOF panel is a promising tool for in-depth profiling of the HGSOC TME.
- The panel enables characterization of individual patient TME profiles.
- Consideration of tissue handling and dissociation methods is crucial for accurate HGSOC TME profiling.
