Targeting FAK‐YAP Signaling Axis Inhibits Tumor Stemness and Overcomes Tumor Resistance to Cuproptosis

ABSTRACT Cuproptosis has recently been identified as a novel metabolic cell death pathway that holds great promise for cancer therapy. However, cuproptosis resistance developed by tumor cells severely impairs its anticancer efficacy. Here, our study uncovers that the FAK‐YAP signaling axis in tumor cells plays important roles in the development of cuproptosis resistance and the promotion of tumor stemness. To therapeutically target this signaling axis, a CRISPR‐cuproptosis synergistic nanoplatform (CPLNP@HA) is elaborately designed for targeted co‐delivery of a Ptk2 (encoding FAK) CRISPR‐Cas9 knockout plasmid to block FAK‐YAP signaling axis and Cu‐elesclomol (Cu‐ES, a cuproptosis inducer) into tumor cells. Mechanistically, CPLNP@HA efficiently silences FAK expression, which subsequently blocks the cuproptosis‐induced activation of the FAK‐YAP signaling cascade. This intervention not only sustains intracellular copper accumulation to reinforce cuproptosis but also reduces tumor stemness, thereby significantly potentiating the therapeutic outcomes of cuproptosis and activating tumoricidal immunity as evidenced by transcriptomic analysis. In combination with immune checkpoint blockade therapy, CPLNP@HA markedly inhibits tumor development upon rechallenge after surgical resection. Our work establishes FAK‐YAP signaling axis as a new therapeutic target to overcome cuproptosis resistance and inhibit tumor stemness, providing a promising combinatorial paradigm to sensitize tumor to cuproptosis and immunotherapy.