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Anthony T Nguyen, Zeljka Smit-McBride, Amar P Patel, David Telander, Lawrence S Morse; Differentially expressedmicroRNAs in serum of patients with wet and dry AMD. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):812. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
This project aims to identify the circulating microRNAs in serum that are differentially expressed in wet and dry age-related macular degeneration (AMD) compared to controls. Specifically we focused on the microRNAs that are involved in stress response and the p53 pathway that plays a role in AMD pathogenesis.
Serum samples of 30 patients from 3 different groups were collected. Group 1 were 10 age-matched normals with no ocular or systemic disease, Group 2 were 10 patients with category 3 high risk dry AMD, and Group 3 were 10 new unilateral exudative AMD patients. RNA was isolated using modified Qiagen microRNeasy procedure, and quantitated on BioAnalyzer’s small RNA chips. Microarray miRNA analysis of serum samples was performed on 4 samples from each group and ran on Affymetrix 2.0 miRNA arrays. GeneSpring software was used for data normalization and ANOVA for statistical analysis. Ingenuity Pathway Analysis (IPA) allowed determination of microRNAs involved in specific cellular pathways. Confirmatory quantitative real-time PCR (qPCR) was used on the select set of microRNAs for the full sample set of each group.
Microarray analysis using GeneSpring software showed several microRNAs that were statistically different (p<.01) between the dry and exudative forms of AMD. MicroRNAs that showed a difference in levels of expression between wet vs dry AMD are hsa-miR-1306 (1.22 vs 1.44, p=.001), hsa-miR-204 (1.60 vs 1.43, p=.001), and hsa-miR-409-3p (1.45 vs 1.95, p=.004). IPA analysis revealed two microRNAs, that are involved in the regulation of angiogenesis and apoptosis, were differentially regulated in wet and dry AMD. Hsa-miR-486-5p was down-regulated in dry AMD compared to controls (17.94 vs 47.76) and hsa-miR-638 which was up regulated in both wet and dry AMD (45.63 and 36.37 vs 22.10). Both of these microRNAs are regulated by p53; MiR-486 has been shown to inhibit silent information regulator 1 (SIRT1), which deacetylates p53, and miR-638 directly targets several apoptosis regulators: p53, PTEN, and BRCA1.
Our study identifies two promising microRNAs that are involved with apoptosis and the p53 pathway that are differentially expressed in dry and wet AMD. These microRNAs warrant further study to determine their contribution to AMD pathology.
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