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Metabolic Profiling of Tumor and Immune Cells Integrating Seahorse and Flow Cytometry
Nienke B Goedhart1,2,3,4, Helga Simon-Molas5,6,7,8
1Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, The Netherlands.
Abstract:
Studying metabolism in the different cellular compartments of the tumor microenvironment (TME) is crucial to identify the specific metabolic signatures that contribute to tumor progression. Additionally, TME-driven metabolic changes can disrupt the essential metabolic processes involved in immune cell function, hindering immunotherapy success. Extracellular flux analysis (Seahorse) is a well-established approach used to characterize cellular metabolism. When combined with flow cytometry-based measurements, it offers a comprehensive view of the metabolic signatures within the different cellular compartments of the TME. Here we provide a detailed description on the procedures of Seahorse analysis and complementary flow cytometry-based metabolic readouts on tumor cells and T cells in the context of cancer, which can also be applied to other physiological and pathological situations.
Insights
Understanding tumor microenvironment (TME) metabolism is key for cancer progression insights. Combining Seahorse analysis with flow cytometry reveals metabolic signatures in TME cells, aiding immunotherapy research.
Area of Science:
- Oncology
- Immunology
- Metabolomics
Background:
- Tumor microenvironment (TME) metabolism significantly influences cancer progression and immune function.
- Metabolic disruptions within the TME can impair anti-tumor immune responses, limiting immunotherapy efficacy.
- Characterizing cellular metabolism in distinct TME compartments is essential for identifying therapeutic targets.
Purpose of the Study:
- To detail Seahorse analysis and flow cytometry methods for assessing TME cellular metabolism.
- To provide a comprehensive approach for analyzing metabolic signatures in tumor cells and T cells.
- To offer a protocol applicable to cancer research and other pathological conditions.
Main Methods:
- Extracellular flux analysis (Seahorse) for cellular metabolic characterization.
- Flow cytometry for complementary metabolic readouts.
- Application of methods to tumor cells and T cells within the TME context.
Main Results:
- Demonstration of Seahorse analysis for TME metabolic profiling.
- Integration of flow cytometry for detailed cellular metabolic assessment.
- Establishment of a combined methodology for comprehensive TME metabolic analysis.
Conclusions:
- The combined Seahorse and flow cytometry approach provides a robust method for dissecting TME metabolic heterogeneity.
- Understanding TME-specific metabolic signatures is critical for developing effective cancer immunotherapies.
- This methodology can be adapted for studying cellular metabolism in various physiological and pathological settings.
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