(+)-Miliusol suppresses the Warburg effect and induces regulated cell death in triple-negative breast cancer through targeting EIF3D and remodeling cancer metabolism

Triple-negative breast cancer (TNBC) exhibits marked molecular heterogeneity, posing ongoing therapeutic challenges. Metabolic reprogramming, particularly through the Warburg effect, offers a promising therapeutic target for TNBC treatment. Data mining and machine learning identified (+)-miliusol as a promising candidate. Its direct target, eukaryotic initiation factor 3D (EIF3D), was validated through mass spectrometry-coupled cellular thermal shift assay (MS-CETSA), a biotinylated probe, and a proteolysis-targeting chimera (PROTAC) approach. EIF3D, an emerging oncoprotein and atypical translation initiation regulator, promotes tumor survival by selectively modulating protein synthesis. (+)-Miliusol demonstrates potent anti-proliferative and anti-migratory activity against TNBC in both in vitro and in vivo . Integrated proteomic and transcriptomic analyses revealed that (+)-Miliusol suppresses TNBC progression through EIF3D-mediated translational regulation. Mechanistically, it disrupts the EIF3D–AlkB homolog 5 (ALKBH5)–glucose transporter type 4 (GLUT4) axis, EIF3D–HIF1 α signaling, and the EIF3D–RuvB like AAA ATPase 1 (RUVBL1)– β -catenin pathway, thereby inhibiting glycolysis and metastasis while inducing ER stress-dependent apoptosis via caspase-12 and JNK activation. Additionally, (+)-miliusol blocks EIF3D–HIF1 α and EIF3D–ALKBH3 interactions, impairing ATAD2/PAK1-regulated Warburg-effect networks and triggering autophagy-associated cell death. (+)-Miliusol induces TNBC cell death by selectively suppressing translation of critical glycolytic and metastatic regulators. These findings establish EIF3D-mediated translational control as a promising therapeutic avenue for TNBC treatment. (+)-Miliusol targets EIF3D to block glycolysis and inactivate multiple oncogenic signals, thereby remodeling cancer metabolism and inducing regulated cell death in TNBC.