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.

解析肿瘤微环境（tumor microenvironment, TME）在肺癌中的作用，对开发新型治疗方案至关重要。T细胞在肿瘤微环境中发挥核心调控作用，亦是多款先进免疫疗法的作用靶点。代谢研究是解析癌细胞与T细胞互作机制的核心所在，因其直接调控肿瘤生长、增殖、免疫应答及药物响应过程。本研究构建了肺癌细胞与两种表型T细胞（初始型与细胞毒性型）的二维体外共培养模型，旨在解析不同细胞群体间代谢协同与竞争的复杂动态过程。本研究采用循环免疫荧光（Cyclic Immunofluorescence, CycIF）技术，以亚细胞分辨率可视化33种代谢酶及调控因子的空间定位特征。相较于初始T细胞群体，细胞毒性T细胞群体的多种T细胞相关标志物表达水平显著升高，且与葡萄糖、氨基酸及脂肪酸代谢相关的代谢谱也更为活跃。研究团队在共培养系统中检测到代谢特征存在显著改变，这表明肺癌细胞与T细胞之间存在动态的代谢互作。本研究揭示了可通过靶向特定代谢通路以增强免疫疗法疗效的潜在靶点，为肿瘤治疗干预提供了全新方向。