AUTHOR=Tian Ye , Hu Xin , Liu Yuan , Wu Wenqi , Yao Yanxin , Liu Huahuan , Wang Wei , Dai Hongji , Huang Yubei , Sun Changyu , Cui Yan , Li Zun , Zhang Xiangnan , Jia Liqing , Wang Fubing , Song Fengju , Chen Kexin , Pan Yuan , Liu Ben 

TITLE=Functional and clinical validation of tsRNA-defined molecular subtypes guides precision therapy in gastric cancer

JOURNAL=Frontiers in Immunology

VOLUME=Volume 16 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2025.1684113

DOI=10.3389/fimmu.2025.1684113

ISSN=1664-3224

ABSTRACT=IntroductionGastric cancer (GC) is a highly heterogeneous malignancy with poor prognosis, underscoring the urgent need for reliable biomarkers to guide precise stratification and therapy. Transfer RNA-derived small RNAs (tsRNAs) have emerged as potential key regulators in cancer, yet their systematic role in defining GC subtypes remains unexplored.MethodsWe profiled tsRNA expression in GC using transcriptomic data from TCGA and GEO databases. Unsupervised consensus clustering identified tsRNA-based subtypes. A prognostic model was constructed using machine learning algorithms and validated across multiple cohorts. The functional role of a key tsRNA, tsRNA-Asp-3-0024, was investigated through Pandora-seq, qRT-PCR, and in vitro and organoid-based assays.ResultsThree distinct tsRNA-mediated subtypes (Stromal_H, Stromal_L, Stromal_M) were identified, exhibiting significant differences in stromal activity, tumor microenvironment, and clinical outcomes. The Stromal_H subtype demonstrated the poorest prognosis, characterized by an immunosuppressive microenvironment and dysregulated DNA repair pathways. A random survival forest (RSF)-based prognostic signature (GCtsRNAscore) effectively stratified patients into high- and low-risk groups, with high-risk patients showing increased sensitivity to targeted therapies (axitinib, bexarotene, dasatinib) and low-risk patients benefiting more from immunotherapy. Furthermore, tsRNA-Asp-3-0024 was significantly upregulated in GC tissues and cell lines, where it promoted proliferation and inhibited apoptosis.DiscussionOur study establishes tsRNAs as powerful biomarkers for molecular subtyping and prognostic prediction in GC. The tsRNA-defined subtypes and GCtsRNAscore model provide a novel framework for personalized treatment strategies. The functional characterization of tsRNA-Asp-3-0024 highlights its potential as both a therapeutic target and a prognostic indicator, paving the way for tsRNA-based precision medicine in GC.