Exosomal microRNA Panels for Detecting Early-Stage Non-Small Cell Lung Cancer

<b>Background</b>: Early diagnosis of lung cancer requires lung nodule biopsies, which can lead to severe complications. This study aimed to identify optimized panels of exosomal microRNAs (miRNAs) for non-invasive diagnosis of early-stage non-small cell lung cancer (NSCLC). <b>Materials and Methods</b>: This study comprised four phases: discovery, validation, optimization, and confirmation. In the discovery phase, next-generation sequencing profiled 2656 exosomal miRNAs in serum samples (<i>n</i> = 76) from patients with benign lung nodules and stage-specific NSCLC. The validation phase used qPCR to analyze selected miRNAs in serum samples (<i>n</i> = 75). The optimization phase employed a self-devised diagnostic platform, the "up-down ratio (UDR)," to identify miRNA panels. The confirmation phase involved miRNA-target gene interaction and enrichment analyses. <b>Results</b>: The discovery phase identified 15 candidate miRNAs, of which six were validated by qPCR: miR-1976, miR-150-5p, miR-301b-3p, miR-369-3p, miR-497-5p, and miR-610. UDR platform identified a panel of four miRNAs optimized for early detection of NSCLC with ROC over 0.93. Bioinformatics analysis revealed 20 target genes, with <i>VEGFA</i>, <i>BCL2</i>, and <i>PTEN</i> showing strong interactions with the miRNAs, particularly with miR-150-5p, miR-205-5p, miR-1976, miR-301b-3p, and miR-497-5p. <b>Conclusions</b>: This four-phase study suggests that exosomal miRNA panels have potential diagnostic value for early-stage lung cancer. The UDR platform enabled the selection of a four-miRNA panel (miR-150-5p, miR-301b-3p, miR-369-3p, and miR-497-5p), with bioinformatics analyses providing supportive evidence.