Deciphering spatial immunometabolism in a 2D co-culture of murine T cells and lung cancer cells using multiplexed imaging tools

Understanding the role of the tumor microenvironment (TME) in lung cancer is crucial for developing new treatments. T cells play a pivotal role within the TME, and are a target of multiple advanced immunotherapies. Metabolism lies at the core of studying the interaction between cancer and T cells because it governs tumor growth, proliferation, immune response, and drug response. Herein, we developed a 2D in-vitro co-culture model with lung cancer cells and several phenotypes of T cells (naïve and cytotoxic) to decipher the intricate dynamics of the metabolic cooperation and competition between the cellular populations. Cyclic Immunofluorescence (CycIF) was used to visualize the spatial localization of 33 metabolic enzymes and regulators at a sub-cellular resolution. The cytotoxic T-cell population showed higher expression in several T-cell related markers, and elevated metabolic profiles associated with glucose, amino acid, and fatty acid metabolism compared to the naïve T-cell population. Significant alterations in metabolic signatures were detected within the co-culture systems. These changes suggest a dynamic metabolic crosstalk between lung cancer cells and T cells. This study sheds light on specific metabolic pathways that can be targeted to enhance the efficacy of immunotherapy, offering new avenues for therapeutic intervention.