SLC25A4 high Hepatocytes Enhance Lifestyle Intervention Response in MASLD by Recruiting Eating Macrophages

ABSTRACT While lifestyle interventions (LSI) are the cornerstone of metabolic dysfunction‐associated steatotic liver disease (MASLD) management, the molecular mechanisms driving the histological and metabolic regression in LSI‐responsive patients remain unclear. C57BL/6N mice were hydrodynamically injected with Slc25a4 and Sleeping Beauty transposon . Single‐cell RNA sequencing (scRNA‐seq) profiled the hepatic immune landscape. Metabolomics and co‐culture assays analyzed intercellular crosstalk. Responsive MASLD patients had high SLC25A4 expression, correlating with exercise‐induced hepatic steatosis improvement. Slc25a4 overexpression augmented exercise effects. scRNA‐seq identified a distinct population of Clec4f + Timd4 + P2ry12 + macrophages, defined as lipid‐eating macrophages, in livers of Slc25a4 ‐overexpressing mice. Macrophage depletion reinstated hepatosteatosis in high‐fat, high‐fructose, and high‐cholesterol diet‐fed exercised mice with Slc25a4 overexpression. Mechanistically, SLC25A4 mediates mitochondrial ATP exchange and selective ATP efflux. Ectopically overexpressed Slc25a4 activates Pannexin 1 to release ATP, which recruits eating macrophages via P2ry12, clearing lipid‐laden hepatocytes. The GDF15‐PPARα axis between macrophages and hepatocytes was activated. Macrophage Gdf15 knockdown abrogated Slc25a4 ‐mediated enhanced hepatic fatty acid oxidation (FAO), and hepatic PPARα knockdown abolished this effect as well. Slc25a4 orchestrates an LSI‐responsive hepatocyte–macrophage axis: Pannexin 1‐mediated ATP release recruits lipid‐eating macrophages, whose GDF15 activates PPARα‐driven FAO, establishing Slc25a4 as a therapeutic target to restore exercise response in MASLD.