Hsp60 lactylation promotes mitochondrial dysfunction and trophoblast apoptosis in preeclampsia

Preeclampsia is characterized by placental hypoxia and metabolic reprogramming toward glycolysis, leading to increased lactate production and protein lactylation. This study investigated the role of heat shock protein 60 (Hsp60) lactylation in preeclampsia pathophysiology. Using placental tissue microarrays and HTR-8/SVneo trophoblast cells, we found that Hsp60 undergoes aberrant lactylation in preeclamptic placentas, particularly under hypoxic conditions. Through in silico prediction and site-directed mutagenesis, we identified K469 and K473 as the primary lactylation sites on Hsp60. Functional studies revealed that Hsp60 lactylation promotes mitochondrial fission by modulating Drp1 phosphorylation, increases mitochondrial ROS production, and sensitizes trophoblasts to apoptosis. Expression of the delactylation-mimetic K469R/K473R mutant significantly attenuated hypoxia-induced mitochondrial fragmentation, preserved mitochondrial membrane potential, reduced cytochrome c release and caspase-3 activation, and maintained trophoblast invasive capacity. Furthermore, we demonstrated that Hsp60 lactylation is regulated by histone deacetylases and p300 acetyltransferase. These findings identify Hsp60 lactylation as a novel mechanism linking metabolic adaptation to mitochondrial dysfunction in preeclampsia. Targeting Hsp60 lactylation or its regulatory enzymes may represent a potential therapeutic strategy for preventing trophoblast dysfunction and improving outcomes in preeclampsia.