Thermodynamic Genome-Scale Metabolic Modeling of Metallodrug Resistance in Colorectal Cancer

SIMPLE SUMMARY: Cancer, but also its treatment, can lead to a reprogramming of cellular metabolism. These changes are observable in metabolite abundances, which can be unbiasedly measured via mass spectrometry metabolomics. However, even when the metabolome changes strongly, a (mechanistic) interpretation is difficult as metabolite levels do not necessarily directly correspond to pathway activities. Here we measure the changes of the cellular metabolome in colorectal cancer cell lines sensitive and resistant to the ruthenium-based drug BOLD-100/KP1339 and the platinum-based drug oxaliplatin. We map these changes onto a cancer-specific genome-scale metabolic model, which allows us not only to compute intracellular flux distributions, but also to disentangle drug-specific effects from growth differences from differences in metabolic adaptations due to resistance. Specifically, we find that resistance to BOLD-100/KP1339 induces more extensive reprogramming than oxaliplatin, especially with respect to fatty acid and amino acid metabolism. ABSTRACT: Background: Mass spectrometry-based metabolomics approaches provide an immense opportunity to enhance our understanding of the mechanisms that underpin the cellular reprogramming of cancers. Accurate comparative metabolic profiling of heterogeneous conditions, however, is still a challenge. Methods: Measuring both intracellular and extracellular metabolite concentrations, we constrain four instances of a thermodynamic genome-scale metabolic model of the HCT116 colorectal carcinoma cell line to compare the metabolic flux profiles of cells that are either sensitive or resistant to ruthenium- or platinum-based treatments with BOLD-100/KP1339 and oxaliplatin, respectively. Results: Normalizing according to growth rate and normalizing resistant cells according to their respective sensitive controls, we are able to dissect metabolic responses specific to the drug and to the resistance states. We find the normalization steps to be crucial in the interpretation of the metabolomics data and show that the metabolic reprogramming in resistant cells is limited to a select number of pathways. Conclusions: Here, we elucidate the key importance of normalization steps in the interpretation of metabolomics data, allowing us to uncover drug-specific metabolic reprogramming during acquired metal-drug resistance.