In conclusion, the loss of ATAD3A triggered accumulated FC and TG, blocked autophagosome degradation and effective degradation of damaged mitochondria with impaired mitochondrial respiration, and induced significantly higher liver cell death when challenged with FC overload, revealing the essential role of ATAD3A in NAFLD through the regulation of lipid homeostasis and mitochondrial health. Here, ATAD3A is linked to metabolic dysfunction-associated steatotic liver disease.