Abstract
Purpose
miRNAs can regulate inflammatory pathways. The purpose of this work was to determine if inflammatory-related tear film miRNAs are associated with extracellular vesicles (EVs) in human non-Sjögren’s Syndrome dry eye disease (DED) participants.
Methods
Five DED and 5 non-DED human participants were recruited. Tears samples were collected by washing the ocular surface of both eyes with phosphate buffered saline, pooling samples from the right and left eyes, and purifying EVs from the samples with a polyethylene glycol (PEG) 8000 precipitation procedure. Samples were directly analyzed via ELISA or transmission electron microscopy (TEM), or RNA was isolated first from the EVs and evaluated with RNA-Seq.
Results
EVs were identified in the tear film of both groups using TEM and ELISA. Following EV purification and RNA isolation, RNA-Seq determined that there were 126 EV miRNAs differentially expressed between the two groups when comparing their RNA cargoes. Ingenuity Pathways Analysis found 9 upregulated miRNAs that were associated with inflammation (miR-127-5p, miR-1273h-3p, miR-1288-5p, miR-130b-5p, miR-139-3p, miR-1910-5p, miR-203b-5p, miR-22-5p, and miR-4632-3p; all p < 0.049; fold regulation range = 1.43–1.67).
Conclusion
This study determined that EVs are present in the tear film and that tear EVs contain miRNAs that may be associated with DED inflammatory pathways.
Disclosure statement
Dr. Andrew D. Pucker has received financial and/or equipment research support from Art Optical Contact Lenses Inc., Alcon, Bausch + Lomb, Contamac, CooperVision, EyeGate Pharmaceuticals Inc., EpiTech, Euclid Systems, Kala, Optikal Care Inc, PentaVision, and WebMD-Jobson Medical Information in the past three years. Dr. Pucker is also currently an employee of Lexitas Pharma Services, though he was solely employed by the University of Alabama at Birmingham during the conduct of this work.
Dr. Cameron Postnikoff is currently employed by CooperVision; however, he was not affiliated with this company while conducting this work.
Dr. Jason J. Nichols in 2019 and 2020 has received honoraria from Paragon Vision Sciences and Coopervision. He has also received research funding from Alcon, Bruder, Johnson and Johnson Vision, and Mallinckrodt over the last 3 years. Also, Dr. Kelly Nichols is the spouse of Dr. Jason Nichols, extending her declarations to him. In the past 12 months, Dr. Kelly Nichols has consulted for and received honorarium from: Bruder, Dompe, Kala, Novartis/Shire (Medical Exchange International), Osmotica, Oyster Point, Sight Sciences, Tear Film Innovations/Alcon/Acquiom, Thea, Tarsus, and TopiVert. She has received research funding from: Allergan, Kala, and Tear Science.
Dr. William Ngo is currently an employee at the Centre for Ocular Research & Education (CORE); however, he was not during the conduct of this work. Over the past three years, members of CORE have received funding from the following companies: Alcon, Allergan, Allied Innovations, Brien Holden Vision Institute, CooperVision, GL Chemtec, I-Med Pharma, Johnson & Johnson Vision, Lubris, Menicon, Nature’s Way, Novartis, Ophtecs, Oté Pharma, PS Therapy, Santen, Shire, SightGlass Vision, and Visioneering. He has received consulting fees from Alcon.
Data availability statement
The authors confirm that the data supporting the results of this study are available within this article and its associated supplementary files (Supplemental Data 1, 2, and 3).