Ultrasound-Visualized Cuproptosis in Glioblastoma via Endogenous Copper Sequestration and Nitric Oxide-Gas Controlled Release Using a Programmable Nanoliposomal Platform

Cuproptosis, a copper-dependent cell death, emerges as a potential anticancer strategy but still faces challenges of systemic toxicity from exogenous copper supplementation, tumor adaptation via glutathione (GSH)-mediated detoxification, and compensatory copper-efflux upregulation. These limitations impede mitochondrial respiratory dysfunction and proteotoxic stress that are essential for cuproptosis, highlighting the demand for tumor-specific copper metabolic modulation. Here, we engineer multifunctional nanoliposomes (DSF/S1P/ISDN-Lipos) that hijack endogenous copper transport for spatially controlled tumor-specific cuproptosis induction while enabling real-time therapeutic monitoring via gas enhanced ultrasonography. Modularly assembled from tumor-targeting sphingosine-1-phosphate (S1P), GSH-responsive nitric oxide (NO) prodrug isosorbide dinitrate (ISDN), and copper-chelator disulfiram (DSF), this DSF/S1P/ISDN-Lipos first facilitates blood-brain tumor barrier traversal and glioblastoma-specific accumulation. Then, intratumorally GSH converts DSF to dithiocarbamate (DTC), chelating endogenous copper into Cu(DTC)₂ complexes on the liposome surface. Following internalization, coreleased Cu(DTC)₂ and ISDN-derived NO deplete GSH while suppressing ATP7B efflux pumps, amplifying copper overload to trigger lipoylated protein aggregation and Fe-S cluster degradation. Notably, NO-generated ultrasound contrast enables spatiotemporal mapping of copper transport dynamics. In vivo, DSF/S1P/ISDN-Lipos demonstrated favorable biosafety and significantly suppressed orthotopic glioblastoma growth. This work presents a theranostic approach for metal homeostasis regulation, where gas therapy synergizes with endogenous metallo-reprogramming to overcome adaptive resistance.