32 cases of peripheral blood small RNA sequencing in humans

Immunochemotherapy has become the standard first-line treatment for advanced non-small cell lung cancer (NSCLC), yet response rates remain limited. Identifying reliable non-invasive biomarkers is crucial for predicting therapeutic benefit and enabling personalized treatment strategies.In this prospective study, we enrolled 32 patients with unresectable locally advanced or metastatic NSCLC receiving tislelizumab combined with carboplatin and paclitaxel. Patients were classified into clinical benefit (CB) and non-clinical benefit (non-CB) groups based on RECIST 1.1 criteria after two treatment cycles. Serum exosomes were isolated via ultracentrifugation before treatment initiation. Small RNA sequencing was performed to profile exosomal miRNA expression, with differential expression validated by quantitative RT-PCR. Target gene prediction and functional enrichment analyses (GO, KEGG, Disease Ontology, Reactome) were conducted for differentially expressed miRNAs using clusterProfiler. ROC curve analysis was conducted to evaluate the predictive performance of candidate miRNAs.We identified 19 differentially expressed exosomal miRNAs between CB and non-CB patients. Seven miRNAs were significantly upregulated in the CB group and validated by RT-qPCR. Among these, miR-192-5p demonstrated the highest predictive accuracy with an AUC of 0.77 (95% CI: 0.543-0.917). Functional enrichment analysis revealed that target genes of these miRNAs were involved in key signaling pathways including MAPK and Ras signaling, as well as biological processes related to apoptosis and cell cycle regulation. Disease Ontology analysis indicated significant association with lung cancer pathogenesis, while Reactome pathway analysis showed enrichment in the RHO GTPase cycle and PI3K/AKT signaling cascades.This study identifies a panel of serum exosomal miRNAs, particularly miR-192-5p, as promising non-invasive biomarkers for predicting response to immunochemotherapy in advanced NSCLC. These findings support the potential clinical application of liquid biopsy-based miRNA profiling to guide treatment selection and improve patient stratification.