Abstract Background: Lung cancer remains the leading cause of cancer-related mortality worldwide, and recurrence frequently occurs even after curative-intent therapy. Reliable tools for early detection, prognosis prediction, and recurrence monitoring are critically needed to improve outcomes. Methods: For early detection of lung cancer, a multicenter case-control study was conducted using urine samples from 278 patients with lung cancer (LC; 50% early-stage) and 213 participants without cancer (NC) collected at four institutions in Japan. Participants were randomly assigned to a training set (208 LC and 159 NC) and a test set (70 LC and 54 NC). Urinary extracellular vesicles-derived microRNAs (EV-miRNAs) were profiled by small RNA sequencing, and machine learning models were developed to evaluate diagnostic performance. For prognosis prediction, urinary miRNA profiles and four-year recurrence-free survival (RFS) were analyzed. Additionally, paired preoperative and postoperative urine samples from 100 surgically treated patients were analyzed, and a subset of patients also provided samples after recurrence to explore temporal miRNA changes during follow-up. Results: The urinary EV-miRNA classifier demonstrated high diagnostic accuracy, with AUCs of 0.942 (95% CI, 0.918-0.966) in the training and 0.941 (95% CI, 0.899-0.983) in the test sets. Sensitivity and specificity for early-stage LC were 82.0% (95% CI, 73.3%-88.3%) and 92.5% (95% CI, 87.3%-95.6%) in the training set, and 88.2% (95% CI, 73.4%-95.3%) and 87.0% (95% CI, 75.6%-93.6%) in the test set, respectively. Eleven miRNAs were significantly associated with RFS, and a prognostic three-miRNA panel (hsa-miR-181a-5p, hsa-miR-185-5p, and hsa-miR-943) was constructed via fivefold cross-validation, yielding a hazard ratio of 8.3 (95% CI, 1.9-37.0). Furthermore, twelve miRNAs decreased after surgery and re-elevated after recurrence in exploratory analyses, indicating preliminary potential for postoperative monitoring. Conclusions: This urinary EV-miRNA-based assay enables highly sensitive, noninvasive early detection of LC and demonstrates potential for prognostic assessment and recurrence monitoring. These findings suggest that a urinary EV-miRNA single-assay concept could provide a multifaceted approach for lung cancer management, supporting early detection, risk stratification, and postoperative surveillance. Citation Format: Yoriko Ando, Ayu Kiritani, Shohei Mori, Takashi Nojiri, Junko Watanabe, Hiroki Yamaguchi, Mika Mizunuma, Yuki Ichikawa, Yasutaka Kato, Jun Araya, Takashi Ohtsuka, Masahiko Higashiyama, Yu Fujita. Urinary extracellular vesicle microRNA profiling enables noninvasive early detection and prognostic stratification of lung cancer abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 1369.
Ando et al. (Fri,) studied this question.