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miRNA Profile in Three Different Normal Human Ocular Tissues by miRNA-Seq
Michelle Drewry1, Inas Helwa1, R Rand Allingham2
1Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, United States.
Investigative Ophthalmology & Visual Science
|July 15, 2016
Summary
This study profiled microRNA (miRNA) expression in human eye tissues, identifying novel miRNAs and their gene targets. This provides a foundation for understanding how miRNA changes contribute to eye diseases like glaucoma and keratoconus.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) play crucial roles in cellular processes and have been implicated in various eye diseases.
- Understanding miRNA expression in healthy ocular tissues is essential for identifying disease-associated alterations.
Purpose of the Study:
- To profile microRNA (miRNA) expression in normal human ciliary body (CB), cornea, and trabecular meshwork (TM) using miRNA sequencing (miRNA-Seq).
- To establish a baseline for miRNA function and involvement in ocular pathologies.
- To identify potential miRNA biomarkers for eye conditions.
Main Methods:
- RNA extraction from human CB, cornea, and TM samples.
- Next-generation sequencing (miRNA-Seq) for comprehensive miRNA profiling.
- Bioinformatic analysis using Bowtie and miRBase for read alignment and identification.
- miRNA target interaction analysis using miRTarBase.
- Validation of selected miRNA expression using droplet digital PCR (ddPCR).
Main Results:
- Identified 378 collectively expressed miRNAs across the three ocular tissues.
- The 11 most abundant miRNAs constituted 80% of total normalized reads.
- Discovered uniquely expressed miRNAs, with five sharing 18 validated gene targets.
- Generated a miRNA profile linked to genes associated with keratoconus and glaucoma.
- Validated the expression of five key miRNAs using ddPCR.
Conclusions:
- First comprehensive miRNA expression profile of human CB, cornea, and TM using miRNA-Seq.
- Identified numerous novel miRNAs in ocular tissues.
- Establishing baseline miRNA expression in healthy ocular tissues can elucidate disease-specific changes in conditions like glaucoma and keratoconus.
