Multistage Responsive NanoCRISPR Unleashes Cascade Amplified Endogenous Apoptosis and Inhibits Epithelial-Mesenchymal Transition Simultaneously for Suppressing Tumor Growth and Metastasis

Targeting apoptotic pathways holds promise for cancer treatment; however, single-pathway approaches often struggle to overcome apoptosis resistance and metastasis. Overexpression of Inhibitor of Apoptosis Proteins (IAPs), particularly Survivin, is critically linked to these therapeutic challenges. Herein, a multistage-responsive nanoCRISPR (MIRV) system targeting the IAPs member Survivin in the nucleus and mitochondria in the cytoplasm was developed to suppress tumor growth and metastasis via cascade-amplified endogenous apoptosis and epithelial-mesenchymal transition (EMT) inhibition. By leveraging the tumor-specific targeting, enzyme-triggered penetrating and deshelling, and reactive oxygen species (ROS)-responsive capabilities, MIRV precisely delivers the Survivin-targeting CRISPR/Cas9 system and the pro-apoptotic peptide D(KLAKLAK)2 to the nucleus and cytoplasm of tumor cells, respectively. Survivin depletion-triggered endogenous apoptosis and EMT inhibition, cooperating with peptide-induced mitochondrial dysfunction, enabled MIRV to markedly suppress subcutaneous tumor growth and peritoneal metastasis with minimal side effects. Taken together, the MIRV system provides an effective strategy for the simultaneous induction of apoptosis and suppression of EMT in antitumor and antimetastatic therapies.