In Situ Generation of Pyroptosis Inducer Mediated by Intracellular Labile Copper Pool for Safe and Robust Antitumor Immunotherapy

Pyroptosis has garnered increasing interest in the realm of cancer immunotherapy. Utilizing reactive oxygen species (ROS) to trigger oxidative stress is considered an effective strategy for promoting pyroptosis. However, existing catalytic nanoparticles used as pyroptosis inducers contain heavy metals, which inevitably cause potential side effects on normal tissues due to their high toxicity and off-target effects. Herein, a labile copper pool-mediated in situ pyroptosis inducer was designed and developed using a hydrogen-bonded organic framework (HOF)-based nanoplatform to achieve safe and robust antitumor immunotherapy. The nanoplatform could target mitochondria and elevate labile Cu²⁺ levels in cells, implementing the in situ synthesis of a pyroptosis inducer through the formation of catalytic nanoparticles with peroxidase (POD) and superoxide dismutase (SOD)-mimicking activities. Our results confirmed that the nanoplatform could generate high levels of ROS, resulting in pyroptotic cell death. When combined with antiprogrammed death receptor 1 therapy (αPD-1), the pyroptosis inducer exhibited excellent antitumor capacity in tumor models. Meanwhile, it exhibited minimal toxicity to healthy tissues due to the low intracellular copper concentration in normal cells. Overall, our work provides potential for the development of efficient and safe antitumor immunotherapy.