Delivery of a ZBP1 agonist enhances radiotherapy-induced antitumour immunity in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) shows limited responsiveness to immune checkpoint inhibitors (ICIs), largely because many tumors remain poorly inflamed and refractory to effective antitumour immunity. Here, we identify Z -DNA-binding protein 1 (ZBP1) as an immune-associated determinant of therapeutic responsiveness in HCC and explore its potential as a target for radio-immunotherapy sensitization. Integrative analysis of necroptosis-related genes and immunotherapy-response datasets revealed that high ZBP1 expression was associated with favorable survival, enhanced antigen-presentation signatures, increased immune-cell infiltration, and improved immunotherapy benefit. Functionally, basal ZBP1 expression alone had limited effects on tumor-cell behavior, whereas pharmacological activation of ZBP1 markedly enhanced therapeutic responsiveness. Mechanistically, ZBP1 activation promoted necroptosis-associated signaling and amplified IFN and STING-related innate immune activation. Radiotherapy further upregulated ZBP1, providing a mechanistic basis for combination treatment. To enable tumor-directed delivery, we developed a glypican-3 (GPC3)-targeted liposomal nanoplatform co-loaded with the ZBP1 agonist CBL0137 and gold nanoparticles (CBL-Au@Lip). CBL-Au@Lip exhibited stable physicochemical properties, acceptable biosafety, and enhanced tumor-associated accumulation. In vitro, CBL-Au@Lip cooperated with radiotherapy to suppress malignant phenotypes and strengthen immune-related signaling. In orthotopic HCC models, triple therapy comprising radiotherapy, anti-PD-L1 antibody (aPD-L1), and CBL-Au@Lip achieved superior tumor control, increased intratumoral T-cell infiltration and cytotoxic activation, and induced durable antitumour immune memory. Importantly, this therapeutic benefit was retained in a fibrosis-associated HCC model, although no obvious antifibrotic effect was observed. Together, these findings identify targeted ZBP1 activation as a promising strategy to enhance radio-immunotherapy and support further translational development of CBL-Au@Lip-based combination therapy in HCC. The GPC3-targeted CBL-Au@Lip nanoparticle platform eradicates HCC by eliciting tumor cell necroptosis and activating a potent T-cell-mediated immune response. The CBL-Au@Lip nanoparticle is synthesized via thin-film hydration using DPPC, cholesterol, CBL0137, and a G12 peptide ligand, followed by rotary evaporation and hydration with Au nanoparticles for intravenous delivery; upon tumor accumulation and GPC3 targeting, it releases CBL0137 to activate the cGAS-STING pathway, inducing type I interferon signaling and promoting CD4+ T cell-mediated antitumor immunity.