Nanoshield‐Assisted Viral Gene Therapy with Induction of Non‐Apoptotic Cell Death and Durable Antitumor Immunity

Non-apoptotic cell death have emerged as promising strategies to overcome apoptotic resistance in cancer therapy. We suggest a hybrid gene delivery platform integrating adeno-associated virus (AAV)-mediated expression of receptor-interacting kinase 3(RIPK3) with manganese dioxide-polyethyleneimine (MnO₂-PEI) to induce necroptosis and immunogenic cell death (ICD), thereby remodeling the tumor microenvironment and enhancing antitumor immunity. This platform combines high transduction efficiency with the tumor-accumulation ability and immunostimulatory potential of non-viral carriers. The MnO₂-PEI nanosheets shields AAV from immune and hepatic clearance, thus enhancing tumor accumulation. This addresses a key limitation of naked AAV delivery. Simultaneously, the AAV payload offsets non-viral systems' low gene delivery efficiency. The platform induces robust damage-associated molecular patterns (DAMP) and tumor antigen release, thereby promoting dendritic cell maturation and cytotoxic T cell infiltration. Furthermore, Mn²⁺-induced reactive oxygen species (ROS) amplify ferroptosis and, in conjunction with RIPK3-mediated necroptosis, remodel the immunosuppressive tumor microenvironment by promoting M1 macrophage polarization and a Th1-type immune response. In tumor re-challenge models, AAV/MnO₂-PEI-treated mice exhibited durable antitumor immunity, thereby highlighting the potential of platform to establish long-term immune memory. This hybrid delivery system provides a potent strategy for synergistic cancer immunotherapy, effectively overcoming the limitations of both viral and non-viral vectors.