Self‐Activated Nitric Oxide Nanogenerators Induce Cuproptosis via Regulating the Endogenous Copper Metabolism for Anti‐PD‐L1 Blockade Therapy

ABSTRACT

Low anti‐PD‐L1 therapy response is mainly caused by poor tumor immunogenicity and insufficient PD‐L1 expression. Cuproptosis, a newly identified copper (Cu)‐dependent cell death, can simultaneously enhance immunogenicity and upregulate PD‐L1, but whether nitric oxide (NO) initiates cuproptosis remains unclear. Herein, we reveal a “NO‐induced cuproptosis” strategy: nanomaterial‐generated NO induces cuproptosis via inhibiting copper chaperone (CCS) expression to trigger endogenous Cu ⁺ release. To optimize this mechanism, we design a non‐copper, self‐activated NO nanogenerator (AZORHP) composed of arginine (Arg)‐stabilized zinc peroxide (AZO) for generating NO via self‐supplied Arg and hydrogen peroxide (H ₂ O ₂ ) reaction, siRNA for silencing the copper‐chelating protein (metallothionein 2A, MT2A), and hyaluronic acid‐polypeptides (HP) decorated with anti‐PD‐L1 peptide and matrix metalloproteinase‐2 (MMP‐2)‐cleavable linkers. AZORHP has been verified to integrate NO‐mediated CCS downregulation to release copper and simultaneously blocking copper chelation via MT2A silencing and copper efflux via ATPase copper transporting β (ATP7B) suppression caused by mitochondrial damage, thereby establishing a dual regulatory network of “copper release–copper retention” that amplifies cuproptosis. This amplified cuproptosis provokes immunogenicity and PD‐L1 upregulation, thereby enhancing the response rate of anti‐PD‐L1 immunotherapy together with MMP‐2‐responsive anti‐PD‐L1 peptide. Therefore, our findings provide a novel concept of NO‐induced cuproptosis via regulating endogenous copper metabolism for immunotherapy.