Language

English

Publication Date

4-1-2026

Journal

The Ocular Surface

DOI

10.1016/j.jtos.2026.01.011

PMID

41638553

PMCID

PMC13050525

PubMedCentral® Posted Date

4-6-2026

PubMedCentral® Full Text Version

Author MSS

Abstract

Purpose: To generate a comprehensive profile of microRNAs (miRNAs) present in human tear fluid using next-generation sequencing (NGS), establish a reference miRNome for healthy human tear fluid, and investigate whether miRNA expression varies by sex, race, or age.

Methods: Tear samples were collected from 32 adults using Schirmer strips. RNA was isolated using the miRNeasy Serum/Plasma Kit, and cDNA libraries were prepared using the QIAseq miRNA Library Kit. Barcoded libraries were sequenced on the NovaSeq 6000 platform. Bioinformatic analyses included adapter trimming, alignment to miRBase, normalization with DESeq2, and functional annotation using multiMiR and clusterProfiler. Differential expression analysis was performed to evaluate variability across demographic groups.

Results: We identified 318 miRNAs constitutively expressed in tear fluid, 246 of which were detected in all subjects. The most abundant miRNAs included hsa-let-7a-5p, hsa-let-7f-5p, hsa-let-7b-5p, hsa-let-7g-5p, and hsa-let-7c-5p. Prominent miRNA families included let-7 (13 members), miR-30 (10), miR-8 (10), miR-10 (8), and miR-17 (8). Functional enrichment analyses indicated that tear miRNAs may regulate pathways involved in transcriptional control, cytoskeletal organization, MAPK signaling, and neurodegenerative processes. Samples processed in two batches several months apart showed strong concordance, confirming methodological reproducibility. Two miRNAs, hsa-miR-375-3p and hsa-miR-708-5p, were significantly increased in White subjects, whereas hsa-miR-106b-3p and hsa-miR-205-3p showed a nonsignificant increase in females.

Conclusions: This study defines a high-resolution reference profile of the human tear fluid miRNome, demonstrating its stability, diversity, and functional relevance. These data establish a robust baseline for future studies investigating disease-associated alterations in tear miRNAs. Observed demographic associations remain exploratory and require larger, adequately powered cohorts.

Keywords

Humans, Female, MicroRNAs, Tears, Male, Adult, High-Throughput Nucleotide Sequencing, Middle Aged, Gene Expression Profiling, Sequence Analysis, RNA, Aged, Young Adult, Tear fluid, Schirmer strip, microRNA, Biomarkers, Small RNA sequencing

Published Open-Access

yes

Download

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.