HIF-2α drives an intrinsic vulnerability to ferroptosis in clear cell renal cell carcinoma

SUMMARY Kidney cancers are characterized by extensive metabolic reprogramming and resistance to a broad range of anti-cancer therapies. By interrogating the Cancer Therapeutics Response Portal compound sensitivity dataset, we show that cells of clear-cell renal cell carcinoma (ccRCC) possess a lineage-specific vulnerability to ferroptosis that can be exploited by inhibiting glutathione peroxidase 4 (GPX4). Using genome-wide CRISPR screening and lipidomic profiling, we reveal that this vulnerability is driven by the HIF-2α–HILPDA pathway by inducing a polyunsaturated fatty acyl (PUFA)-lipid-enriched cell state that is dependent on GPX4 for survival and susceptible to ferroptosis. This cell state is developmentally primed by the HNF-1β–1-Acylglycerol-3-Phosphate O-Acyltransferase 3 (AGPAT3) axis in the renal lineage. In addition to PUFA metabolism, ferroptosis is facilitated by a phospholipid flippase TMEM30A involved in membrane topology. Our study uncovers an oncogenesis-associated vulnerability, delineates the underlying mechanisms and suggests targeting GPX4 to induce ferroptosis as a therapeutic opportunity in ccRCC. HIGHLIGHTS ccRCC cells exhibit strong susceptibility to GPX4 inhibition-induced ferroptosis The GPX4-dependent and ferroptosis-susceptible state in ccRCC is associated with PUFA-lipid abundance The HIF-2α–HILPDA axis promotes the selective deposition of PUFA-lipids and ferroptosis susceptibility AGPAT3 selectively synthesizes PUFA-phospholipids and primes renal cells for ferroptosis