Bladder cancer is a common malignancy in urology, marked by a high recurrence rate and limited treatment strategies. This highlights the need for novel biomarkers and therapeutic targets. The RUVBL1/2 complex, consisting of RUVBL1 and RUVBL2 and functioning as a AAA+ ATPase, plays a crucial role in various cellular mechanisms. Dysregulation of RUVBL1/2 has been implicated in cancer, with studies showing that both genetic and pharmacological inhibition can suppress tumorigenic properties. However, the specific contribution of the RUVBL1/2 complex to bladder cancer remains largely unexplored.

A genome-wide CRISPR/Cas9 screening using the Brunello library in J82 and RT4 cell lines identified RUVBL1 and RUVBL2 as druggable vulnerabilities in bladder cancer. To investigate the role of the RUVBL1/2 complex in tumor progression, CRISPR/Cas9-mediated depletion and pharmacological inhibition were employed. Functional analyses included proliferation, BrdU incorporation, cell cycle and apoptosis, 2D clonogenic and 3D soft agar anchorage-independent growth assays, migration and invasion, and DNA Damage and Replication Stress assays. Additionally, the effects of pharmacological inhibition were assessed across these assays, emphasizing its therapeutic potential.

Genetic depletion or pharmacological inhibition of the RUVBL1/2 complex using the selective inhibitor CB-6644 significantly impaired cell proliferation, survival, and 3D anchorage-independent growth, leading to cell cycle arrest and increased apoptosis. Mechanistically, these effects were accompanied by a reduction in migration and invasion capacities, supported by elevated DNA damage, replication stress, and alterations in key molecular markers, further highlighting the therapeutic potential of targeting RUVBL1/2 in bladder cancer.

Our findings indicate a significant reliance of bladder cancer on the RUVBL1/2 complex, emphasizing its potential as both a biomarker and a therapeutic target. The functional and mechanistic roles of RUVBL1/2 in tumor progression warrant further investigation, particularly in the context of its cellular and molecular interactions. Future studies exploring these pathways will provide deeper insights into the contribution of RUVBL1/2 to bladder cancer pathogenesis and may aid in the development of targeted therapeutic strategies.