Abstract 4297: Interrogating metabolic heterogeneity and cooperation in lung cancer collective cell invasion

Abstract Tumor heterogeneity contributes to tumor progression and a variable response to treatment. We have previously shown that tumor heterogeneity - including metabolic, transcriptional and mutational - underlies non-small cell lung cancer (NSCLC) collective invasion. Using stable subpopulations derived with our Spatiotemporal Cellular and Genomic Analysis (SaGA) technique, we found that two distinct subpopulations of the H1299 NSCLC line cooperate during collective tumor invasion in vitro; while H1299 pack leaders are heavily dependent on oxidative phosphorylation, H1299 pack followers are more reliant on glycolysis. To further dissect how intratumoral heterogeneity enables cancer progression to metastasis, we incorporate high resolution imaging, stable fluorescent labeling, gene editing, and in vitro tumor invasion models to interrogate mechanisms of metabolic cooperation between heterogeneous NSCLC subpopulations. We hypothesize that cooperation and communication between these distinct metabolic subpopulation drives metastatic tumor progression. To determine mechanisms of metabolic cooperativity between subpopulations, we stably transduced our subpopulations to fluorescently label either the mitochondria (through TOM20 labeling) or a key enzyme in glycolysis (through glucose 6-phosphate isomerase GPI labeling), cultured labeled subpopulations with unlabeled subpopulations, and observed metabolic changes and exchange via time-lapse, fluorescence microscopy and flow cytometry in 2D culture conditions. Next steps include visualizing the metabolic cooperation that occurs in conditions mimicking the tumor microenvironment (i.e., 3D invasion assays) using super-resolution microscopy. Taken together, we have developed a method of temporally visualizing metabolic exchange with high spatial resolution between NSCLC subpopulations to investigate the underlying mechanisms enabling successful cell invasion in a variety of tumor microenvironments. Thus far, we have discovered multiple broad mechanisms of exchange including tunneling nanotubes and extracellular vesicles, both of which are inducible with certain stressors common to the tumor microenvironment. Citation Format: Isaac E. Robinson, Veronika Matsuk, Mala Shanmugam, Janna K. Mouw, Adam I. Marcus. Interrogating metabolic heterogeneity and cooperation in lung cancer collective cell invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4297.