<p>Colorectal cancer (CRC) remains one of the most common and deadly malignancies, with limited effective treatment options. Cuproptosis is a novel regulated cell death triggered by mitochondrial copper overload (Cu²⁺) and has been implicated in several cancers. However, the mechanisms governing cuproptosis in CRC are largely unknown. In this study, we investigated the role of ubiquitin-specific proteases (USPs), key regulators of protein stability, in modulating cuproptosis in CRC. Through integrative analysis of publicly available CRC transcriptomic datasets, we identified oncogenic USPs potentially involved in cuproptosis regulation. Among them, USP11 emerged as significantly overexpressed in CRC tissues, and its elevated expression was correlated with poorer patient survival. We next examined the functional role of USP11 in two CRC cell lines and patient-derived CRC organoids. Cells were treated with elesclomol and CuCl₂, a combination known to induce cuproptosis. Functional assays revealed that USP11 promotes cell viability, colony formation, survival, and migration/invasion and suppresses cuproptosis in response to treatment. Mechanistic studies showed that USP11 directly binds to and deubiquitinates ISCU, a scaffold protein essential for iron–sulfur cluster assembly and known to bind Cu²⁺. This deubiquitination stabilizes ISCU, thereby preventing its proteasomal degradation. Rescue experiments further confirmed that ISCU is a key downstream effector mediating USP11’s cuproptosis resistance effect. Finally, in patient-derived CRC organoid models, knockdown of USP11 significantly enhanced cuproptosis upon elesclomol/CuCl₂ treatment, validating our in vitro findings. Collectively, our work identifies USP11 as a novel inhibitor of cuproptosis in CRC, acting through the stabilization of ISCU. This newly uncovered USP11–ISCU axis contributes to tumor resistance against cuproptotic stress and highlights a potential therapeutic target for sensitizing CRC to copper-induced cell death.</p>