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bioRxiv · 10.1101/2024.10.14.618123

Pyruvate dehydrogenase kinase 1 controls triacylglycerol hydrolysis in cardiomyocytes

Atser, M. G.·Wenyonu, C. D.·Rowe, E. M.·Leung, C. L. K.·Cen, H. H.·Queathem, E. D.·Liu, L. T.·Moravcova, R.·Rogalski, J.·Perrin, D.·Crawford, P.·Foster, L. J.·Alcazar, A.·Johnson, J. D.
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Abstract

Pyruvate dehydrogenase kinase (PDK) 1 is one of four isozymes that inhibit the oxidative decarboxylation of pyruvate to acetyl-CoA via pyruvate dehydrogenase. PDK activity is elevated in fasting or starvation conditions to conserve carbohydrate reserves. PDK has also been shown to increase mitochondrial fatty acid utilization. In cardiomyocytes, metabolic flexibility is crucial for the fulfillment of high energy requirements. The PDK1 isoform is abundant in cardiomyocytes, but its specific contribution to cardiomyocyte metabolism is unclear. Here we show that PDK1 regulates cardiomyocyte fuel preference by mediating triacylglycerol turnover in differentiated H9c2 myoblasts using lentiviral shRNA to knockdown Pdk1. Somewhat surprisingly, PDK1 loss did not affect overall PDH activity, basal glycolysis, or glucose oxidation revealed by oxygen consumption rate experiments and 13C6 glucose labelling. On the other hand, we observed decreased triacylglycerol turnover in H9c2 cells with PDK1 knockdown, which was accompanied by decreased mitochondrial fatty acid utilization following nutrient deprivation. 13C16 palmitate tracing of uniformly labelled acyl chains revealed minimal acyl chain shuffling within triacylglycerol, indicating that the triacylglycerol hydrolysis, and not re-esterification, was dysfunctional in PDK1 suppressed cells. Importantly, PDK1 loss did not significantly impact the cellular lipidome or triacylglycerol accumulation following palmitic acid treatment, suggesting that effects of PDK1 on lipid metabolism were specific to the nutrient-deprived state. We validated that PDK1 loss decreased triacylglycerol turnover in Pdk1 knockout mice. Together, these findings implicate a novel role for PDK1 in lipid metabolism in cardiomyocytes, independent of its canonical roles in glucose metabolism.

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Atser, M. G., Wenyonu, C. D., Rowe, E. M., Leung, C. L. K., Cen, H. H., Queathem, E. D., Liu, L. T., Moravcova, R., Rogalski, J., Perrin, D., Crawford, P., Foster, L. J., Alcazar, A., Johnson, J. D.. 2024-10-14. Pyruvate dehydrogenase kinase 1 controls triacylglycerol hydrolysis in cardiomyocytes. https://doi.org/10.1101/2024.10.14.618123

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