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

Abstract 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 (MnO 2 ‐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 MnO 2 –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/MnO 2 –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.