Upregulation of glycolysis and downregulation of mitochondrial oxidative phosphorylation, termed as the Warburg effect, is characteristic of tumor cells. Restriction of pyruvate flux into the mitochondrial matrix is one of the major mechanisms underlying this phenomenon. Warburg metabolism is beneficial for rapidly proliferating cells; however, the metabolic consequences of activation of mitochondrial respiration in such cells remain unclear. Here we created a chemogenetic instrument, Grubraw, that generates pyruvate from ᴅ-alanine directly in the mitochondrial matrix, bypassing restricted pyruvate influx. In cancer cells, Grubraw-produced mitochondrial pyruvate increased oxidative metabolism. Chemogenetic activation of mitochondria slowed the growth rate of human melanoma xenografts. Moreover, activation of cancer cell mitochondria suppressed the transcription of genes that control cell-cycle progression, cell proliferation and DNA replication. Surprisingly, we found that cancer cells with Warburg metabolism control carbon flux by disposing of excess mitochondrial pyruvate produced by Grubraw and exporting it out of the cell. The chemogenetic tool Grubraw activates mitochondria by generating pyruvate in the mitochondrial matrix. This triggers a genetic switch that slows down tumor cell proliferation. Tumor cells counteract this by exporting excess pyruvate outward.
Pyruvate links glycolysis to mitochondrial oxidative metabolism, while the Warburg effect diverts it to lactate. A xenotopic chemogenetic tool based on bacterial d-alanine dehydrogenase generates pyruvate in the mitochondrial matrix independently of glycolysis, resulting in enhanced respiration and pyruvate export.
