Universität zu Köln

Toyono, Tetsuya, Usui, Tomohiko, Villarreal, Guadalupe, Jr., Kallay, Laura, Matthaei, Mario, Vianna, Lucas M. M., Zhu, Angela Y., Kuroda, Masahiko, Amano, Shiro and Jun, Albert S. (2016). MicroRNA-29b Overexpression Decreases Extracellular Matrix mRNA and Protein Production in Human Corneal Endothelial Cells. Cornea, 35 (11). S. 1466 - 1471. PHILADELPHIA: LIPPINCOTT WILLIAMS & WILKINS. ISSN 1536-4798

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Abstract

Purpose:MicroRNAs are small noncoding RNAs that regulate gene expression at the posttranscriptional level. We reported that levels of microRNA (miR)-29 family are decreased in corneas of patients with Fuchs endothelial corneal dystrophy (FECD). The miR-29 family regulates the production of extracellular matrix (ECM) proteins. Accumulation of ECM proteins in Descemet membrane is an important pathologic change in FECD. In this study, we transfected miR-29b into human corneal endothelial cells and tissues and evaluated ECM protein expression levels.Methods:An immortalized Fuchs human corneal endothelial cell line (iFECD) was established by infection of corneal endothelial cells from patients with FECD with hTERT lentivirus. MiR-29b was transfected into iFECD, and the expression levels of ECMs collagen type 1 alpha 1 (COL1A1), collagen type 4 alpha 1 (COL4A1), and laminin gamma 1 (LAMC1) were evaluated with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and Western blot. Expression level of LAMC1 protein in miR-29b-transfected donor corneal endothelium was also evaluated by Western blot.Results:Compared with control, miR-29b expression level after transfection of iFECD was increased to 335.6% (91.0%), and ECM expression levels were significantly decreased. Compared with control, qRT-PCR demonstrated reduction of ECM to the following levels: COL1A1: 1.9% (+/- 0.4%); COL4A1: 7.1% (+/- 1.7%); and LAMC1: 21.5% (+/- 2.7%). Western blot showed reduced protein expression: COL1A1: 4.8% (+/- 3.2%); COL4A1: 42.5% (+/- 25.0%); and LAMC1: 44.8% (+/- 3.1%). In miR-29b-transfected corneal tissue, LAMC1 protein expression level was decreased to 14.4% (+/- 20.5%).Conclusions:Overexpression of miR-29b decreased ECM protein production in human corneal endothelial cells. Thus, miR-29 replacement therapy might be a new treatment strategy for FECD aimed at reducing pathologic production of ECM proteins in Descemet membrane.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Toyono, Tetsuya UNSPECIFIED UNSPECIFIED UNSPECIFIED Usui, Tomohiko UNSPECIFIED UNSPECIFIED UNSPECIFIED Villarreal, Guadalupe, Jr. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kallay, Laura UNSPECIFIED UNSPECIFIED UNSPECIFIED Matthaei, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED Vianna, Lucas M. M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Zhu, Angela Y. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuroda, Masahiko UNSPECIFIED UNSPECIFIED UNSPECIFIED Amano, Shiro UNSPECIFIED UNSPECIFIED UNSPECIFIED Jun, Albert S. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-257267
DOI:	10.1097/ICO.0000000000000954
Journal or Publication Title:	Cornea
Volume:	35
Number:	11
Page Range:	S. 1466 - 1471
Date:	2016
Publisher:	LIPPINCOTT WILLIAMS & WILKINS
Place of Publication:	PHILADELPHIA
ISSN:	1536-4798
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language OXIDATIVE STRESS; PENETRATING KERATOPLASTY; FUCHS DYSTROPHY; EXPRESSION; COLLAGEN; MIR-29B; COL8A2; DIFFERENTIATION; L450W; DICER Multiple languages Ophthalmology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/25726