HIF-1-alpha links mitochondrial perturbation to the dynamic acquisition of breast cancer tumorigenicity

Up-regulation of hypoxia-inducible factor-1 beta (HIF-1 beta), even in normoxia, is a common feature of solid malignancies. However, the mechanisms of increased HIF-1 beta abundance, and its role in regulating breast cancer plasticity are not fully understood. We have previously demonstrated that dimethyl-2-ketoglutarate (DKG), a widely used cell membrane-permeable beta-ketoglutarate (beta-KG) analogue, transiently stabilizes HIF-1 beta by inhibiting prolyl hydroxylase 2. Here, we report that breast cancer tumorigenicity can be acquired through prolonged treatment with DKG. Our results indicate that, in response to prolonged DKG treatment, mitochondrial respiration becomes uncoupled, leading to the accumulation of succinate and fumarate in breast cancer cells. Further, we found that an early increase in the oxygen flux rate was accompanied by a delayed enhancement of glycolysis. Together, our results indicate that these events trigger a dynamic enrichment for cells with pluripotent/stem-like cell markers and tumorsphere-forming capacity. Moreover, DKG-mediated metabolic reprogramming results in HIF-1 beta induction and reductive carboxylation pathway activation. Both HIF-1 beta accumulation and the tumor-promoting metabolic state are required for DKG-promoted tumor repopulation capacity in vivo. Our data suggest that mitochondrial adaptation to DKG elevates the ratio of succinate or fumarate to beta-KG, which in turn stabilizes HIF-1 beta and reprograms breast cancer cells into a stem-like state. Therefore, our results demonstrate that metabolic regulation, with succinate and/or fumarate accumulation, governs the dynamic transition of breast cancer tumorigenic states and we suggest that HIF-1 beta is indispensable for breast cancer tumorigenicity.