USP7 inhibitor P5091 enhances the antitumor efficacy of vemurafenib in BRAFV600E-mutant thyroid cancer via ferroptosis.

Resistance to the BRAF inhibitor vemurafenib (PLX4032) limits its efficacy in thyroid cancer. Ubiquitin-specific peptidase 7 (USP7), a key regulator of oncogenic signaling, and USP7 inhibitor may help overcome drug resistance. This study investigated the combined efficacy of PLX4032 and the USP7 inhibitor P5091 in BRAFV600E-mutant thyroid cancer. Bioinformatics showed that USP7 and integrin subunit beta 3 (ITGB3), a MAPK/PI3K pathway gene, may jointly mediate resistance. In thyroid cancer cell lines, the combination treatment significantly reduced viability, proliferation, colony formation, migration, and invasion versus monotherapy. Moreover, the combination treatment can reduce viability and induce cell death in thyroid cancer organoids. Given USP7's role in oxidative stress and ferroptosis, we examined its involvement and found that P5091 induced ferroptosis via reactive oxygen species (ROS) elevation, glutathione peroxidase 4 (GPX4) downregulation, and elevated lipid peroxidation. These findings demonstrate that USP7 inhibition by P5091 enhances PLX4032 efficacy by promoting tumor suppression and ferroptosis in BRAFV600E-mutant thyroid cancer, offering a promising strategy to overcome resistance.