Electron transport inhibition increases cellular dependence on purine transport and salvage

Wu, Z.; Bezwada, D.; Harris, R. C.; Pan, C.; Nguyen, P. T.; Faubert, B.; Cai, L.; Cai, F.; Vu, H. S.; Chen, H.; Martin-Sandoval, M.; Do, D.; Gu, W.; Zhang, Y.; Ko, B.; Brooks, B.; Kelekar, S.; Zacharias, L. G.; Oaxaca, K. C.; Mathews, T. P.; Garcia-Bermudez, J.; Ni, M.; DeBerardinis, R. J.

doi:10.1101/2023.05.11.540429

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bioRxiv · 10.1101/2023.05.11.540429

Electron transport inhibition increases cellular dependence on purine transport and salvage

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Abstract

Cancer cells reprogram their metabolism to support cell growth and proliferation in harsh environments. While many studies have documented the importance of mitochondrial oxidative phosphorylation (OXPHOS) in tumor growth, some cancer cells experience conditions of reduced OXPHOS in vivo and induce alternative metabolic pathways to compensate. To assess how human cells respond to mitochondrial dysfunction, we performed metabolomics in fibroblasts and plasma from patients with inborn errors of mitochondrial metabolism, and in cancer cells subjected to inhibition of the electron transport chain (ETC). All these analyses revealed extensive perturbations in purine-related metabolites; in non-small cell lung cancer (NSCLC) cells, ETC blockade led to purine metabolite accumulation arising from a reduced cytosolic NAD+/NADH ratio (NADH reductive stress). Stable isotope tracing demonstrated that ETC deficiency suppressed de novo purine nucleotide synthesis while enhancing purine salvage. Analysis of NSCLC patients infused with [U-13C]glucose revealed that tumors with markers of low oxidative mitochondrial metabolism exhibited high expression of the purine salvage enzyme HPRT1 and abundant levels of the HPRT1 product inosine monophosphate (IMP). ETC blockade also induced production of ribose-5 phosphate (R5P) by the pentose phosphate pathway (PPP) and import of purine nucleobases. Blocking either HPRT1 or nucleoside transporters sensitized cancer cells to ETC inhibition, and overexpressing nucleoside transporters was sufficient to drive growth of NSCLC xenografts. Collectively, this study mechanistically delineates how cells compensate for suppressed purine metabolism in response to ETC blockade, and uncovers a new metabolic vulnerability in tumors experiencing NADH excess.

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Wu, Z., Bezwada, D., Harris, R. C., Pan, C., Nguyen, P. T., Faubert, B., Cai, L., Cai, F., Vu, H. S., Chen, H., Martin-Sandoval, M., Do, D., Gu, W., Zhang, Y., Ko, B., Brooks, B., Kelekar, S., Zacharias, L. G., Oaxaca, K. C., Mathews, T. P., Garcia-Bermudez, J., Ni, M., DeBerardinis, R. J.. 2023-05-11. Electron transport inhibition increases cellular dependence on purine transport and salvage. https://doi.org/10.1101/2023.05.11.540429

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