eCite Digital Repository

Mutational analysis of MIR184 in sporadic keratoconus and myopia

Citation

Willoughby, CE and Lechner, J and Bae, HA and Guduric-Fuchs, J and Rice, A and Govindarajan, G and Siddiqui, S and Farraj, LA and Yip, SP and Yap, M and Das, M and Souzeau, E and Coster, D and Mills, RA and Lindsay, R and Phillips, T and Mitchell, P and Ali, M and Inglehearn, CF and Sundaresan, P and Craig, JE and Simpson, DA and Burdon, KP, Mutational analysis of MIR184 in sporadic keratoconus and myopia, Investigative Ophthalmology and Visual Science (Iovs), 54, (8) pp. 5266-5272. ISSN 0146-0404 (2013) [Refereed Article]


Preview
PDF
Restricted - Request a copy
587Kb
  

Copyright Statement

Copyright 2013 Association for Research in Vision and Opthamology

DOI: doi:10.1167/iovs.13-12035

Abstract

PURPOSE. A mutation miR-184({thorn}57C>T) in the seed region of miR-184 (encoded by MIR184 [MIM*613146]) results in familial severe keratoconus combined with early-onset anterior polar cataract and endothelial dystrophy, iris hypoplasia, congenital cataract, and stromal thinning (EDICT) syndrome (MIM#614303). In order to investigate the phenotypic spectrum resulting from MIR184 mutation, MIR184 was sequenced in a keratoconus cohort of mixed ethnicity and a Chinese axial myopia cohort. METHODS. Sequencing of MIR184 was performed in 780 unrelated keratoconus patients and 96 unrelated Han southern Chinese subjects with axial myopia. Effects of identified mutations on RNA secondary structure were predicted computationally using mFold and RNAFold algorithms. MIR184 amplicons from patients harboring mutations were cloned and transfected into human embryonic kidney 293T (HEK293T) cells, and mature mutant miR184 expression was analyzed by stem-loop real-time quantitative PCR (RT-qPCR). RESULTS. Two novel heterozygous substitution mutations in MIR184 were identified in the two patients with isolated keratoconus: miR-184({thorn}8C>A) and miR-184({thorn}3A>G). Computational modeling predicted that these mutations would alter the miR-184 stem-loop stability and secondary structure. Ex vivo miR-184 expression analysis demonstrated that miR184({thorn}8C>A) almost completely repressed the expression of miR-184 (P = 0.022), and miR-184({thorn}3A>G) reduced the expression of miR-184 by approximately 40% (P = 0.002). There was no significant association of rs41280052, which lies within the stem-loop of miR184, with keratoconus. No MIR184 mutations were detected in the axial myopia cohort. CONCLUSIONS. Two novel heterozygous substitution mutations in MIR184 were identified in two patients with isolated keratoconus: miR-184({thorn}8C>A) and miR-184({thorn}3A>G). Mutations in MIR184 are a rare cause of keratoconus and were found in 2 of 780 (0.25%) cases.

Item Details

Item Type:Refereed Article
Keywords:Corneal dystrophies; Hereditary; Hsa-miR-184; Keratoconus; Mir184; Myopia
Research Division:Medical and Health Sciences
Research Group:Ophthalmology and Optometry
Research Field:Ophthalmology
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Hearing, Vision, Speech and Their Disorders
Author:Burdon, KP (Associate Professor Kathryn Burdon)
ID Code:90082
Year Published:2013
Web of Science® Times Cited:16
Deposited By:Menzies Institute for Medical Research
Deposited On:2014-03-26
Last Modified:2014-04-29
Downloads:0

Repository Staff Only: item control page