Ceramides have been shown to elicit the tissue damage that underlies a broad spectrum of cardiometabolic disorders, including the liver damage that drives metabolic-associated steatotic liver disease. Using a mouse model of hepatocyte-specific ceramide overproduction, we show a dramatic, sexually dimorphic phenotype of ceramide-driven hepatocyte death and dysfunction. In male mice, hepatocyte-specific overexpression of a functional heterotrimer of serine palmitoyl transferase (SPT) elicits a marked increase in liver ceramides coupled to acute mortality and liver injury. Remarkably, female mice were refractory to this intervention. This model presents us with a unique opportunity to uncover the precise mechanism(s) of ceramide-induced hepatocyte injury and death, while gaining insight into a potential protective role of the female biological sex. Using discovery methods like proteome integral solubility assays and CRISPRi screening, we have identified novel ceramide effectors that contribute to mitochondrial damage and hepatocyte death. These studies provide important insight into the molecular machinery that drives hepatocyte damage, which could have important implications in a broad array of disease processes.