Phenotypic Characterization by Mass Cytometry of the Microenvironment in Ovarian Cancer and Impact of Tumor Dissociation Methods

• Published in: Cancers Vol. 13; no. 4; p. 755
• Main Authors: Anandan, Shamundeeswari, Thomsen, Liv Cecilie V, Gullaksen, Stein-Erik, Abdelaal, Tamim, Kleinmanns, Katrin, Skavland, Jørn, Bredholt, Geir, Gjertsen, Bjørn Tore, McCormack, Emmet, Bjørge, Line
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 11.02.2021; MDPI
• Subjects: Antibodies; Antigens; Cell suspensions; Cytometry; Embedding; Fibroblasts; Gene expression; Lymphocytes; Malignancy; Methods; Ovarian cancer; Patients; Stem cell transplantation; Stem cells; Stochasticity; T cell receptors; Tumor microenvironment; Tumors; high-grade serous ovarian cancer (HGSOC); Cytometry by Time-of-Flight (CyTOF); tumor microenvironment (TME); cell expression profile; tumor dissociation; single-cell mass cytometry; characterization
• ISSN: 2072-6694; 2072-6694
• DOI: 10.3390/cancers13040755

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 ( = 1), stromal ( = 2), and tumor ( = 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.