Differentially expressed miRNA profiles of serum-derived exosomes in patients with sudden sensorineural hearing loss

Juhong Zhang; Haizhu Ma; Guijun Yang; Jing Ke; Wenfang Sun; Li Yang; Shaojing Kuang; Hai Li; Wei Yuan

doi:10.3389/fneur.2023.1177988

Differentially expressed miRNA profiles of serum-derived exosomes in patients with sudden sensorineural hearing loss

Front Neurol. 2023 Jun 2:14:1177988. doi: 10.3389/fneur.2023.1177988. eCollection 2023.

Authors

Juhong Zhang^{1

2}, Haizhu Ma¹, Guijun Yang¹, Jing Ke^{1

2}, Wenfang Sun¹, Li Yang¹, Shaojing Kuang¹, Hai Li³, Wei Yuan^{1

2}

Affiliations

¹ Department of Otorhinolaryngology Head and Neck Surgery, Chongqing General Hospital, Chongqing, China.
² School of Basic Medicine, Chongqing Medical University, Chongqing, China.
³ Department of Otorhinolaryngology Head and Neck Surgery, Xuanhan County People's Hospital, Dazhou, Sichuan, China.

Abstract

Objectives: This study aimed to compare the expressed microRNA (miRNA) profiles of serum-derived exosomes of patients with sudden sensorineural hearing loss (SSNHL) and normal hearing controls to identify exosomal miRNAs that may be associated with SSNHL or serve as biomarkers for SSNHL.

Methods: Peripheral venous blood of patients with SSNHL and healthy controls was collected to isolate exosomes. Nanoparticle tracking analysis, transmission electron microscopy, and Western blotting were used to identify the isolated exosomes, after which total RNA was extracted and used for miRNA transcriptome sequencing. Differentially expressed miRNAs (DE-miRNAs) were identified based on the thresholds of P < 0.05 and |log₂fold change| > 1 and subjected to functional analyses. Finally, four exosomal DE-miRNAs, including PC-5p-38556_39, PC-5p-29163_54, PC-5p-31742_49, and hsa-miR-93-3p_R+1, were chosen for validation using quantitative real-time polymerase chain reaction (RT-qPCR).

Results: Exosomes were isolated from serum and identified based on particle size, morphological examination, and expression of exosome-marker proteins. A total of 18 exosomal DE-miRNAs, including three upregulated and 15 downregulated miRNAs, were found in SSNHL cases. Gene ontology (GO) functional annotation analysis revealed that target genes in the top 20 terms were mainly related to "protein binding," "metal ion binding," "ATP binding," and "intracellular signal transduction." Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that these target genes were functionally enriched in the "Ras," "Hippo," "cGMP-PKG," and "AMPK signaling pathways." The expression levels of PC-5p-38556_39 and PC-5p-29163_54 were significantly downregulated and that of miR-93-3p_R+1 was highly upregulated in SSNHL. Consequently, the consistency rate between sequencing and RT-qPCR was 75% and sequencing results were highly reliable.

Conclusion: This study identified 18 exosomal DE-miRNAs, including PC-5p-38556_39, PC-5p-29163_54, and miR-93-3p, which may be closely related to SSNHL pathogenesis or serve as biomarkers for SSNHL.

Keywords: biomarkers; exosomes; miRNA; miRNA transcriptome sequencing; sudden sensorineural hearing loss.

Grants and funding

This study was supported by the Chongqing Science and Health Joint General Project (No. 2022MSXM046), the Medical Science and Technology Innovation Fund of Chongqing People's Hospital (Key Project) (No. Y2020ZDXM01), High-end Medical Talents of Young and Middle-aged People in Chongqing (7th, Organization Department of Chongqing Municipal Committee of CPC, Chongqing Municipal Health Commission), the Young and Middle-aged Medical Excellence Team in Chongqing (the first batch) Chongqing Science, and the Health Joint Major Medical Project (No. 2022DBXM006).