Exercise Metabolic Memory Halts Pathological Cardiac Hypertrophy via PDK4

Methods

We employed PET/CT to assess cardiac glucose uptake and bulk RNA sequencing to profile myocardial gene expression in sedentary (Sed) and EHP mice. Genetic manipulation of Pdk4 was achieved via adeno-associated virus mediated overexpression and tamoxifen-inducible, cardiac-specific knockout (Pdk4-CKO). Pressure overload was induced by transverse aortic constriction (TAC) in Pdk4-CKO and control (Myh6-CreER, MCM) mice.

Epigenetic regulation of Pdk4 by EHP was investigated using pyrosequencing, SELECT-qPCR and dual-luciferase assays. Untargeted metabolomics and molecular docking, molecular dynamics simulation, and CETSA were performed on heart tissues and neonatal rat cardiomyocytes/fibroblasts to identify key metabolites and their mechanisms of action. Results: EHP conferred sustained myocardial glucose preference even after regression of physiological hypertrophy, mediated through METTL3-dependent m6A RNA methylation that suppressed Pdk4 expression.

Pdk4 overexpression abolished EHP-mediated cardioprotection, whereas Pdk4 deletion enhanced cardiac function and attenuated fibrosis under pressure overload. Metabolomic profiling identified arachidonic acid-derived metabolites 5-KETE, 12-keto-leukotriene B4, and 20-hydroxy-leukotriene B4 as novel inhibitors of hypertrophy and fibrosis. These metabolites attenuated cardiomyocyte hypertrophy and fibroblast trans-differentiation through inhibition of the ERK2/MAPK1 pathway.

Conclusions: This study establishes a unified mechanism by which EHP induces metabolic memory through RNA methylation-dependent suppression of Pdk4, leading to altered arachidonic acid metabolism and the accumulation of protective lipid mediators. These findings highlight the therapeutic potential of targeting the PDK4–arachidonic acid metabolites axis to mitigate pathological cardiac remodeling.