Cancer-associated fibroblasts drive metabolic heterogeneity in colorectal cancer cells: predictions from metabolic modeling.

KRAS-mutant colorectal cancer (CRC) undergoes metabolic reprogramming that promotes tumor progression and drug resistance. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment (TME), play a pivotal role in modulating these metabolic adaptations in CRC. This study applies flux sampling combined with representation learning and hierarchical clustering to a computational model of central carbon metabolism to understand how CAFs influence KRAS-mutant CRC metabolic reprogramming following targeted enzyme knockdowns. Focusing on 12 key enzymes involved in glycolysis and the pentose phosphate pathway, knockdowns were simulated under normal CRC media and CAF-conditioned media (CCM) conditions. Analysis revealed CCM induces greater metabolic heterogeneity, with knockdown models exhibiting more variable and distinct metabolic states compared to those cultured in normal CRC media, indicating CAF-derived factors diversify the metabolic responses of CRC cells to enzyme perturbations. Pathway-level flux analysis demonstrated media-specific shifts in central carbon metabolism. Predicted biomass flux showed enzyme knockdowns reduced growth across both conditions, but CCM models indicated a protective effect against perturbation. Overall, simulations illustrated CCM enhances the metabolic adaptability of KRAS-mutant CRC cells to perturbations, emphasizing the importance of including TME components in metabolic modeling and therapeutic development and suggesting that targeting tumor-CAF metabolic interactions may improve treatment strategies.