Mechanisms of FOXK1-regulated glycolipid metabolism in mediating TOX-induced histone lactylation to promote CD8⁺ T cell exhaustion in high-grade serous ovarian cancer.

To determine whether FOXK1 induces CD8⁺ T cell exhaustion via histone lactylation in high-grade serous ovarian cancer (HGSOC). Cellular studies utilized qRT-PCR and Western blotting to compare FOXK1 expression in normal ovarian vs. cancer cells. Western blot assessed proteins linked to aerobic glycolysis, lipid metabolism, histone ubiquitination, and epithelial-mesenchymal transition (EMT). Cell migration/invasion were evaluated via scratch and Transwell assays. In vivo, a mouse ovarian cancer model was established. Lactate and lipid levels in supernatants/tissues were measured using Oil Red O and detection kits. TOX and glucose and lipid metabolism regulatory factors were analyzed by H&E staining and immunohistochemistry. The expression levels of immune related factors and the proportion of positive immune detection points in the supernatant of CD8+ T cell culture and tumor tissue were detected by ELISA kits and flow cytometry. Ovarian cancer cells showed elevated FOXK1, glycolysis proteins, lipid regulators, histone ubiquitination, EMT markers, lactate, and lipids compared to normal cells. Tumor tissues exhibited higher glycolysis proteins, lipid regulators, ubiquitination, and lipids than non-cancerous tissue. FOXK1 knockdown in SKOV3 cells reduced lactate, lipids, glucose uptake, glycolysis/lipid proteins, and inhibited proliferation/migration/invasion, while enhancing CD8 + T cell proliferation, immune checkpoint positivity, and immune factors, with decreased apoptosis. In mice, FOXK1 knockdown reduced tumor volume, TOX, glycolipid regulators, ubiquitination, and lipids, but increased immune factors and checkpoint-positive cells in tissues. FOXK1 regulates glycolipid metabolism and TOX histone lactylation, driving CD8 + T cell exhaustion, suggesting novel immunotherapy targets. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-32938-3.