Multi-omics reveals a new mechanism by which GARS1 drives bladder cancer progression through ASNS: purine metabolism reprogramming, mTORC1 activation, and intratumoral microbiota dysbiosis

Bladder cancer (BC), a common urinary system malignancy, and its progression mechanism is complex, involving the activation of multiple signaling pathways. Recent studies have indicated that members of the Aminoacyl-tRNA synthetase (AARS) family play crucial roles in tumor metabolic reprogramming. However, the function and mechanisms of Glycyl-tRNA Synthetase 1(GARS1) in BC remain unclear. In this study, We integrated multi-omics analyses (transcriptomics, metabolomics, 16S microbiomics) with experimental validation to explore GARS1-related molecules and pathways in BC progression. Clinical samples were analyzed via HE staining, immunohistochemistry, and Western blotting to assess GARS1 expression. Si-GARS1 T24 cell lines were constructed using siRNA technology, followed by transcriptomic and metabolomic profiling. In vivo tumor growth was evaluated in BALB/c nude mice, and intratumoral microbial diversity was analyzed via 16S rRNA sequencing. GARS1 expression was significantly higher in bladder cancer tissues than in normal tissues, and elevated GARS1 expression was closely associated with poor patient prognosis. Multi-omics integrated analysis revealed that asparagine synthetase (ASNS) is an interacting protein of GARS1, and knockdown of GARS1 led to significant alterations in purine metabolism-related metabolites. In vivo experiments showed that GARS1 knockdown inhibited tumor growth with a tumor volume inhibition rate of 53.5%. In summary, GARS1 regulates ASNS expression, activates the Mammalian Target of Rapamycin Complex 1 (mTORC1) signaling pathway, disrupts purine metabolism, and impairs tissue microenvironment homeostasis, thereby accelerating bladder cancer progression. This finding not only highlights the important role of GARS1 in bladder cancer but also provides a theoretical basis for developing new therapeutic targets.