NR4A1 Modulates Glioblastoma Sensitive to Erastin-Induced Ferroptosis via NCOA4 mediated Autophagy

Autophagy-mediated ferritin degradation (ferritinophagy) is recognized as a critical driver of ferroptosis; however, the molecular circuitry linking selective autophagy to ferroptosis in glioma remains incompletely defined. Here, we demonstrate that the orphan nuclear receptor NR4A1 as an essential orchestrator of autophagy initiation during ferroptosis. Analysis of TCGA data revealed that lower NR4A1 expression correlates with higher glioma grade and worse patient prognosis. Knockdown of NR4A1 confers robust resistance to erastin-induced ferroptosis in human glioma cells.Mechanistically, erastin exposure triggers a rapid NR4A1-dependent cytoplasmic translocation, enabling direct interaction with the cargo receptor NCOA4. This interaction facilitates the autophagic sequestration and lysosomal degradation of ferritin, thereby amplifying the intracellular ferrous iron pool and propagating lipid peroxidation-driven ferroptosis. NR4A1 knockdown disrupts this axis, resulting in ferritin retention, diminished ferrous iron availability, and suppression of reactive oxygen species (ROS) generation. Conversely, Overexpression of NR4A1 up-regulates NCOA4 expression, and accelerates ferritinophagy, culminating in heightened ferroptotic sensitivity. Pharmacological inhibition of autophagy or knockdown of ATG7 substantially mitigates erastin-induced ferroptosis by diminishing NR4A1-mediated accumulation of intracellular ferrous iron and ROS. In vivo orthotopic xenograft models employing U87MG and GL261 glioblastoma cells stably expressing NR4A1-targeting shRNA corroborate these findings: tumors with NR4A1 depletion exhibit diminished autophagic activity, reduced ferritin turnover, and marked resistance to erastin-mediated ferroptosis, leading to accelerated tumor growth. Conversely, pharmacological activation of NR4A1 by the Cytosporone B (Csn-B) restrains malignant progression and significantly prolongs survival in glioma models.Collectively, our data establish NR4A1 as a pivotal molecular switch that couples autophagy induction to ferritinophagy-dependent ferroptosis in glioma cells and position NR4A1 pharmacology as a therapeutic strategy for enhancing ferroptosis in aggressive glioblastoma.