Evolution of Relapse-Proficient Subclones Constrained by Collateral Sensitivity to Oncogene Overdose in Wnt-Driven Mammary Cancer

Targeted cancer therapeutics select for drug-resistant rescue subclones (RSCs), which typically carry rescue mutations that restore oncogenic signaling. Whereas mutations underlying antibiotic resistance frequently burden drug-naive microbes with a fitness cost, it remains unknown whether and how rescue mutations underlying cancer relapse encounter negative selection prior to targeted therapy. Here, using mouse models of reversible, Wnt-driven mammary cancer, we uncovered stringent counter-selection against Wnt signaling overdose during the clonal evolution of RSCs. Analyzing recurrent tumors emerging during simulated targeted therapy (Wnt withdrawal) by multi-region DNA sequencing revealed polyclonal relapses comprised of multiple RSCs, which bear distinct but functionally equivalent rescue mutations that converge on sub-maximal Wnt pathway activation. When superimposed on native (i.e., undrugged) signaling, these rescue mutations faced negative selection, indicating that they burden RSCs with a fitness cost before Wnt withdrawal unmasks their selective advantage. Exploiting collateral sensitivity to oncogene overdose may help eliminate RSCs and prevent cancer relapse. [Display omitted] •Wnt-driven mammary cancers maintain dependence on sub-maximal Wnt signaling•Rescue mutations destined to subvert targeted therapy carry a fitness cost•Oncogene overdose imposes negative selection, leading to turnover of rescue subclones Keller and Gunther show that Wnt-driven mammary cancers challenged with simulated targeted therapy (Wnt withdrawal) undergo clonal evolution, which stringently selects for mutations that restore a “just right” level of oncogenic signaling. Therefore, cancer relapses emerge from rare subclones that are encumbered by an untapped vulnerability to oncogene overdose.