Simple SummaryIron-sulfur clusters (ISCs) play a crucial role in cancer cell survival and growth, and their dysregulation is associated with the development and progression of cancer. This review highlights the link between ISC metabolism and ferroptosis, a new form of regulated cell death induced by iron-dependent accumulation of lethal lipid peroxidation. The review summarizes the current knowledge on the mechanisms of ISC biogenesis, the role of ISC modulation in ferroptosis, and the potential of targeting ISCs for cancer therapy. The review provides insight into new therapeutic strategies for cancer treatment based on regulating ISC metabolism.AbstractIron dysregulation is a hallmark of cancer, characterized by an overexpression of genes involved in iron metabolism and iron-sulfur cluster (ISC) biogenesis. Dysregulated iron homeostasis increases intracellular labile iron, which may lead to the formation of excess cytotoxic radicals and make it vulnerable to various types of regulated cell death, including ferroptosis. The inhibition of ISC synthesis triggers the iron starvation response, increasing lipid peroxidation and ferroptosis in cancer cells treated with oxidative stress-inducing agents. Various methods, such as redox operations, iron chelation, and iron replacement with redox-inert metals, can destabilize or limit ISC formation and function, providing potential therapeutic strategies for cancer treatment. Targeting ISCs to induce ferroptosis represents a promising approach in cancer therapy. This review summarizes the state-of-the-art overview of iron metabolism and ferroptosis in cancer cells, the role of ISC modulation in ferroptosis, and the potential of targeting ISCs for ferroptosis induction in cancer therapy. Further research is necessary to develop and validate these strategies in clinical trials for various cancers, which may ultimately lead to the development of novel and effective treatments for cancer patients.