Metabolic balance in colorectal cancer is maintained by optimal Wnt signaling levels.

Wnt pathways are important for the modulation of tissue homeostasis, and their deregulation is linked to cancer development. Canonical Wnt signaling is hyperactivated in many human colorectal cancers due to genetic alterations of the negative Wnt regulator APC. However, the expression levels of Wnt‐dependent targets vary between tumors, and the mechanisms of carcinogenesis concomitant with this Wnt signaling dosage have not been understood. In this study, we integrate whole‐genome CRISPR/Cas9 screens with large‐scale multi‐omic data to delineate functional subtypes of cancer. We engineer APC loss‐of‐function mutations and thereby hyperactivate Wnt signaling in cells with low endogenous Wnt activity and find that the resulting engineered cells have an unfavorable metabolic equilibrium compared with cells which naturally acquired Wnt hyperactivation. We show that the dosage level of oncogenic Wnt hyperactivation impacts the metabolic equilibrium and the mitochondrial phenotype of a given cell type in a context‐dependent manner. These findings illustrate the impact of context‐dependent genetic interactions on cellular phenotypes of a central cancer driver mutation and expand our understanding of quantitative modulation of oncogenic signaling in tumorigenesis. Synopsis: Whole‐genome CRISPR screens in genome‐engineered colorectal cancer (CRC) cell lines combined with large‐scale multi‐omic data integration reveal the role of diverging patterns of Wnt hyperactivation during the development of distinct CRC subtypes. Genome‐engineering of APC mutations leads to Wnt‐hyperactivation in CRC cell lines with low endogenous Wnt signaling activity.The dosage level of oncogenic Wnt hyperactivation impacts the metabolic equilibrium and mitochondrial phenotypes of tumors.These findings suggest context‐dependent genetic interactions and quantitative modulation of oncogenic signaling during tumorigenesis. [ABSTRACT FROM AUTHOR]