June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
MicroRNA29b over-expression decreases extra-cellular matrix protein secretion in Fuchs endothelial corneal dystrophy endothelial cells.
Author Affiliations & Notes
  • Tetsuya Toyono
    Wilmer Eye Institute, Johns Hopkins, Baltimore, MD
    Ophthalmology, University of Tokyo, Tokyo, Japan
  • Guadalupe Villarreal
    Wilmer Eye Institute, Johns Hopkins, Baltimore, MD
  • Laura Kallay
    Wilmer Eye Institute, Johns Hopkins, Baltimore, MD
  • Tomohiko Usui
    Ophthalmology, University of Tokyo, Tokyo, Japan
  • Shiro Amano
    Inoue Eye Hospital, Tokyo, Japan
  • Albert S Jun
    Wilmer Eye Institute, Johns Hopkins, Baltimore, MD
  • Footnotes
    Commercial Relationships Tetsuya Toyono, None; Guadalupe Villarreal, None; Laura Kallay, None; Tomohiko Usui, None; Shiro Amano, None; Albert Jun, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1175. doi:
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      Tetsuya Toyono, Guadalupe Villarreal, Laura Kallay, Tomohiko Usui, Shiro Amano, Albert S Jun; MicroRNA29b over-expression decreases extra-cellular matrix protein secretion in Fuchs endothelial corneal dystrophy endothelial cells.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1175.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: MicroRNAs are small non-coding RNAs which regulate gene expression at the post-transcriptional level. We have reported that the microRNA (miR)-29 family which regulates extracellular matrix (ECM) expression is decreased in Fuchs endothelial corneal dystrophy (FECD) patient corneas compared to normal corneas. Accumulation of ECM proteins in Descemet membrane is an important pathologic change in FECD. In this study, we transfected miR29b into immortalized human corneal endothelial cells and evaluated ECM protein expression levels.

Methods: An immortalized FECD human corneal endothelial cell line was established by infection of an FECD patient’s corneal endothelial cells with hTERT lentivirus.MiR29b was transfected into human FECD corneal endothelial cells using RNAiMAX (Invitrogen) for 48 hours. Total RNA and protein were extracted. MiR29b expression change was evaluated with qPCR and ECM proteins (COL1A1, COL4A1, LAMC1) levels were evaluated with qPCR and Western blot.

Results: Compared with control, miR29b expression level was increased to 335.6(±91.0)% and ECM protein expression levels were decreased. In qPCR results for miR29b transfected cells vs. controls, COL1A1 was 1.9(±1.4)%, COL4A1 was 7.2(±3.0)% and LAMC1 was 21.5(±7.0)%. In Western blot results for miR29b transfected cells vs. controls, COL4A1 was 42.5(±25.0)% and LAMC1 was 44.8(±3.1)%.

Conclusions: Over-expression of miR29b decreased ECM protein expression in FECD human corneal endothelial cells. Thus, miR29 replacement might be a new treatment strategy for FECD aimed at reducing pathologic production of ECM proteins in Descemet membrane.

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