Pharmacological targeting of cholesterol biosynthesis in cancer: Sterol intermediates, oxysterols, and sterol flux rewiring

• Cholesterol biosynthesis inhibition rewires sterol metabolic flux in cancer cells. • Sterol intermediates and oxysterols act as signaling molecules in tumors. • 5,6-epoxycholesterol metabolism links oxidative stress to tumor regulation. • Biological effects of cholesterogenesis inhibitors depend on enzymatic context. • Identification of new sterols requires synthesis-guided analytical approaches. Cholesterol metabolism has emerged as an important regulator of tumor cell biology. Beyond its structural role in membranes, the cholesterol biosynthetic pathway generates sterol intermediates and oxidized derivatives able to modulate signaling pathways controlling proliferation, differentiation, and survival. Early studies mainly emphasized the requirement of the mevalonate pathway for cholesterol synthesis and for production of non-sterol isoprenoid intermediates involved in protein prenylation, but increasing evidence indicates that the biological effects of inhibiting cholesterogenesis cannot be explained solely by cholesterol depletion. Blocking specific enzymatic steps frequently causes accumulation of sterol precursors that differ structurally from cholesterol and are highly susceptible to oxygenation under conditions of oxidative stress commonly observed in tumor cells, leading to formation of bioactive oxysterols. Among these metabolites, cholesterol-5,6-epoxides represent a central metabolic node linking sterol imbalance, lipid peroxidation, and sterol signaling. Depending on the enzymatic context, these epoxides can be converted into metabolites with tumor-promoting or tumor-suppressive properties, indicating that inhibition of cholesterol biosynthesis redistributes sterol metabolic flux rather than simply reducing cholesterol levels. Identification of sterol metabolites formed after perturbation of cholesterogenesis remains difficult because many derivatives differ only by subtle structural modifications or exist as stereoisomers, often requiring chemical synthesis of reference compounds and dedicated analytical methods. These observations support the concept of sterol flux rewiring, in which the biological outcome of targeting cholesterol biosynthesis depends on the nature of accumulated intermediates, the oxidative environment, and the enzymatic context of the cell, with important implications for pharmacological targeting of sterol metabolism in cancer.