Mutational analysis of MIR184 in sporadic keratoconus and myopia
| Autor: | Layal Abi Farraj, D. J. Coster, Ha Ae Bae, Judith Lechner, Paul Mitchell, Manir Ali, Maurice Yap, Gowthaman Govindarajan, Emmanuelle Souzeau, David Simpson, Richard A. Mills, Manoranjan Das, Colin E. Willoughby, Jasenka Guduric-Fuchs, Tony Phillips, Kathryn P. Burdon, Shea Ping Yip, Jamie E Craig, Salina Siddiqui, Richard G Lindsay, Periasamy Sundaresan, Chris F. Inglehearn, Aine Rice |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Keratoconus genetic structures Genotype Mutant DNA Mutational Analysis Anterior polar cataract medicine.disease_cause Real-Time Polymerase Chain Reaction medicine Myopia Humans Genetics corneal dystrophies Mutation business.industry MIR184 Dystrophy DNA Middle Aged medicine.disease Phenotype eye diseases Pedigree MicroRNAs Real-time polymerase chain reaction Mutation testing Female hsa-miR-184 business hereditary |
| Zdroj: | Lechner, J, Bae, H A, Guduric-Fuchs, J, Rice, A, Govindarajan, G, Siddiqui, S, Abi Farraj, L, Yip, S P, Yap, M, Das, M, Souzeau, E, Coster, D, Mills, R A, Lindsay, R, Phillips, T, Mitchell, P, Ali, M, Inglehearn, C F, Sundaresan, P, Craig, J E, Simpson, D A, Burdon, K P & Willoughby, C E 2013, ' Mutational Analysis of MIR184 in Sporadic Keratoconus and Myopia ', Investigative Opthalmology and Visual Science, vol. 54, no. 8, pp. 5266-72 . https://doi.org/10.1167/iovs.13-12035 |
| ISSN: | 1552-5783 |
| DOI: | 10.1167/iovs.13-12035 |
| Popis: | Author version made available in accordance with the publisher's policy. A mutation miR-184(+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. 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 miR-184 expression was analyzed by stem-loop real-time quantitative PCR (RT-qPCR). Two novel heterozygous substitution mutations in MIR184 were identified in the two patients with isolated keratoconus: miR-184(+8C>A) and miR-184(+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 miR-184(+8C>A) almost completely repressed the expression of miR-184 (P = 0.022), and miR-184(+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 miR-184, with keratoconus. No MIR184 mutations were detected in the axial myopia cohort. Two novel heterozygous substitution mutations in MIR184 were identified in two patients with isolated keratoconus: miR-184(+8C>A) and miR-184(+3A>G). Mutations in MIR184 are a rare cause of keratoconus and were found in 2 of 780 (0.25%) cases. Supported by Fight for Sight (United Kingdom; JL, CEW); The Research and Development Office, Northern Ireland (RRG Grant 4.46; CEW); Biotechnology and Biological Sciences Research Council, United Kingdom (Grant BB/H005498/1; JG-F, DAS); National Health and Medical Research Council of Australia (KPB, JEC); ALCON India (GG, MD, PS); and The Aravind Eye Care System (GG, MD, PS). JL is a Fight for Sight PhD student. |
| Databáze: | OpenAIRE |
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