Sequencing and validation of exosomal miRNAs panel as novel plasma biomarkers for early diagnosis and prognosis prediction in laryngeal cancer

Abstract

Purpose Plasma exosomal miRNAs are important biomarkers for body fluid biopsy. The purpose of this study was to screen and construct plasma exosomal miRNAs panel as biomarkers for diagnosis and prognosis in laryngeal squamous cell carcinoma (LSCC). Methods Plasma exosomal miRNAs from 6 LSCC patients with three typical anatomical sites and 3 normal controls (NC) were analyzed by next-generation sequencing. The obtained aberrant expression profile of exosomal miRNAs were compared with the online databases of LSCC to construct and verify the diagnostic and prognostic panel by machine learning. Also, quantitative real-time polymerase chain reaction (qRT-PCR) was applied to validate the diagnostic efficacy of the screened miRNAs in an independent clinical cohort. Results A plasma exosomal miRNAs panel (consisting of hsa-miR-139-3p, hsa-miR-486-5p, hsa-miR-944, hsa-miR-320b and hsa-miR-455-5p) was successfully constructed for early diagnosis and prognosis of LSCC, and showed good predictive potential with an AUC of 0.782, 1.000, 0.716, and 0.875 by artificial neural network (ANN) panel in the independent datasets. This panel was further validation in an independent cohort consisting of 84 clinical cases (48 LSCC and 36 NC). In the validation cohort, the AUC of 5 individual miRNAs ranged from 0.721 to 0.837. The accuracy was further increased by the logistic model, which further increased the AUC to 0.959 by adjusting for the number of miRNAs. The mRNA-miRNA regulatory network and immune function analysis revealed the possible underlying pathogenesis of LSCC. Conclusions Exosomal miRNAs panel can be promising plasma biomarkers for predicting early diagnosis and prognosis in LSCC.