Ultrasound Activated Hybrid‐Biomimetic Nanocarriers That Combine Tumor‐Confined CRISPR/Cas9 Metabolic Reprogramming and Cuproptosis With Anticancer Macrophage Polarization

ABSTRACT Nanomedicine aims to develop nanocarriers that provide strong cell selectivity and efficient intracellular delivery. Additionally, therapeutic strategies are expanding to include metabolic pathways to trigger apoptosis and reduce tumor growth, especially in cases resistant to conventional chemotherapy. Here, we have created nanocarriers with hybrid‐biomimetic coatings that, upon ultrasound activation, release encapsulated copper‐based metal‐organic frameworks (MOFs) and COP1 gene knockout Cas9 ribonucleoproteins (RNPs). This hybrid‐membrane coating, which combines tumor and immune cell membranes with perfluorocarbons, enhances tumor‐to‐normal cell uptake and allows for controlled release and cytolytic entry of the nanocarrier contents. We observe that the RNPs efficiently knockout the COP1 gene, thereby arresting the cancer cell cycle in the G0/G1 phase and promoting mitochondrial respiration over anaerobic glycolysis. This increased respiration makes cancer cells more susceptible to cuproptosis triggered by the MOFs and decreases tumor lactate levels, preventing lactate‐driven M2 polarization of tumor‐infiltrating macrophages. Furthermore, the nanocarriers' cellular selectivity leaves macrophages unharmed. These effects enable infiltrating macrophages to retain an anticancer M1 polarization and continue to foster a more active immune response. The combination of tumor‐specific genetic metabolic reprogramming and enhanced cuproptosis activity, along with increased immune activity, results in significant tumor growth suppression and improved survival rates.