Translational dysregulation plays a key role in tumour initiation and progression, including in colorectal cancer (CRC). However, the mechanisms underlying translational control and the systematic characterization of translation-dependent vulnerabilities remain poorly understood in CRC.

We profiled 34 colorectal tissue samples to define a comprehensive translatomic landscape of CRC, uncovering a set of unannotated ORFs from coding (both in-frame and out-of-frame) and long noncoding RNAs (lncRNAs). Additionally, we delineated five translatomic clusters that significantly correlated with specific clinical features and common CRC mutations.

Further analysis revealed a functionally coherent network of 449 genes that are exclusively regulated at the translational level and specifically promote CRC progression by modulating cell growth and immune responses. Among these, tumour necrosis factor receptor 2 (TNFR2) stood out as the most promising vulnerability with significant translational activation in CRC. Pharmacological inhibition of TNFR2 suppressed tumorigenesis in both cell-based and organoid models, and was further validated in vivo using patient-derived organoid xenograft (PDOX) models.

Our study establishes a systematic framework bridging unbiased translatome discovery to functional validation, in which we identified and functionally validated the therapeutic vulnerability of translationally activated TNFR2 in CRC.