Suppressor tRNAs restore functionally enhanced tumour suppressor protein to induce tumour regression

Nonsense mutations in tumour suppressor genes (TSGs) drive cancer initiation and progression but remain “undruggable” due to formidable challenges in rescuing the functional impairment of heterogeneous truncated proteins with a single agent. Here we observe that suppressor transfer RNAs (sup-tRNAs) can overcome diverse nonsense mutations in TP53 , restoring full-length p53 expression and function. Sup-tRNAs enable high-fidelity incorporation of optimal amino acids and can further enhance the transcriptional activity of p53 to suppress tumour cell proliferation. Moreover, we employ a computational model to guide sup-tRNA design by in silico saturation mutagenesis for targeted readthrough of different nonsense-mutant TSGs. Remarkably, sup-tRNAs produced p53 variants with enhanced activity relative to restored wild-type p53, reducing cancer cell colony area by up to 82%. In two xenograft models harbouring nonsense mutant TP53 , we observed up to a 74% reduction in tumour volume after lipid nanoparticle (LNP)-based sup-tRNA treatment, attributed to robust recovery of p53 expression and function. Critically, sup-tRNA monotherapy surpassed nab-paclitaxel chemotherapy in efficacy, and their combination induced near-complete tumour regression (a maximum of 93% reduction in tumour volume) without detectable toxicity in multiple mouse tumour models. Our findings highlight the potential of tRNA-based therapy to provide a promising strategy for rescuing nonsense-mutant TP53 .