title: miRNA Profile in Three Different Normal Human Ocular Tissues by miRNA-Seq.
abstract: Because microRNAs (miRNAs) have been associated with eye diseases, our study aims to profile ocular miRNA expression in normal human ciliary body (CB), cornea, and trabecular meshwork (TM) using miRNA-Seq to provide a foundation for better understanding of miRNA function and disease involvement in these tissues.
  
  
  
  Total RNAs were extracted from seven normal human CB, seven cornea, and seven TM samples using mirVana total RNA isolation kit. miRNA-Seq was done with Illumina MiSeq. Bowtie software was used to trim and align generated sequence reads, and only exact matches to mature miRNAs from miRBase were included. The miRTarBase database was used to analyze miRNA target interactions, and the expression of five selected miRNAs was validated using droplet digital PCR (ddPCR).
  
  
  
  Using the miRNA extracted from 21 human samples, we found 378 miRNAs collectively expressed, of which the 11 most abundant miRNAs represented 80% of the total normalized reads. We also identified uniquely expressed miRNAs, of which five share 18 highly validated gene targets, and created a profile of miRNAs known to target genes associated with keratoconus and glaucoma. Using ddPCR, we validated the expression profile of five miRNAs from miRNA-Seq.
  
  
  
  For the first time, we profiled miRNA expression in three human ocular tissues using miRNA-Seq, identifying many miRNAs that had not been previously reported in ocular tissue. Defining the relative expression of miRNAs in nondiseased eye tissues could help uncover changes in miRNA expression that accompany diseases such as glaucoma and keratoconus.
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, United States.
authors.forename: Michelle
authors.initials: M
authors.lastname: Drewry
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, United States.
authors.forename: Inas
authors.initials: I
authors.lastname: Helwa
authors.affiliation: Department of Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States.
authors.forename: R Rand
authors.initials: RR
authors.lastname: Allingham
authors.affiliation: Department of Medicine and Ophthalmology, Duke University Medical Center, Durham, North Carolina, United States.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Hauser
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, United States 4Center for Biotechnology and Genomic Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, United States 5Ja.
authors.forename: Yutao
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000368:Aged; D002924:Ciliary Body / Q000378:metabolism*; D003315:Cornea / Q000378:metabolism*; D005260:Female; D005901:Glaucoma / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007640:Keratoconus / Q000235:genetics / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D008875:Middle Aged; D017423:Sequence Analysis, RNA; D014129:Trabecular Meshwork / Q000378:metabolism*
chemical_list: D035683:MicroRNAs