April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Differential miRNA Expression In Geographic Atrophy Retina And Retinal Pigment Epithelium
Author Affiliations & Notes
  • Philip Ruzycki
    N-NRL, National Eye Institute, NIH, Bethesda, Maryland
  • Matthew Brooks
    N-NRL, National Eye Institute, NIH, Bethesda, Maryland
  • Harsha Rajasimha
    N-NRL, National Eye Institute, NIH, Bethesda, Maryland
  • Sami Dridi
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Hiroki Kaneko
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Bradley Gelfand
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Valeria Tarallo
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Judit Baffi
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Jayakrishna Ambati
    Department of Ophthalmology & Visual Sciences, University of Kentucky, Lexington, Kentucky
  • Anand Swaroop
    N-NRL, National Eye Institute, NIH, Bethesda, Maryland
  • Footnotes
    Commercial Relationships  Philip Ruzycki, None; Matthew Brooks, None; Harsha Rajasimha, None; Sami Dridi, University of Kentucky (P); Hiroki Kaneko, University of Kentucky (P); Bradley Gelfand, University of Kentucky (P); Valeria Tarallo, University of Kentucky (P); Judit Baffi, University of Kentucky (P); Jayakrishna Ambati, University of Kentucky (P); Anand Swaroop, None
  • Footnotes
    Support  NIH Intramural Program, NIH Grant R01EY018836, NIH Grant R21EY019778, Doris Duke Charitable Foundation, Burroughs Wellcome Fund
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3918. doi:https://doi.org/
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      Philip Ruzycki, Matthew Brooks, Harsha Rajasimha, Sami Dridi, Hiroki Kaneko, Bradley Gelfand, Valeria Tarallo, Judit Baffi, Jayakrishna Ambati, Anand Swaroop; Differential miRNA Expression In Geographic Atrophy Retina And Retinal Pigment Epithelium. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3918. doi: https://doi.org/.

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Abstract
 
Purpose:
 

Changes in gene expression have been reported as a consequence of aging and may contribute to the development of many aging-associated diseases. miRNA mediated gene regulation is thus of particular interest in the genesis of age-related macular degeneration. To test the hypothesis that the development of geographic atrophy, the advanced form of atrophic macular degeneration, is associated with differential miRNA expression, we utilized next generation sequencing to determine global profiles of miRNA expression in retinal pigment epithelium (RPE) and neural retina samples from 4 age matched control and 4 affected individuals.

 
Methods:
 

Total RNA was purified from isolated human retina and RPE. Small RNA was apportioned by flashPAGE. Libraries were prepared using the Illumina small RNA v1.5 protocol and size selected for adaptor ligated 22bp miRNA. Completed libraries were loaded on the Illumina Cluster Station and sequenced on the Illumina GAIIx with 36 cycles of base incorporation. Image analysis and base calling was performed using the Illumina Pipeline. Reads were trimmed of adaptor sequences and all 15-35 bp sequences were mapped to the human genome and miRbase. Biological replicates were pooled and tag counts assessed for each miRNA.

 
Results:
 

The sequence data yielded about 82 million reads for affected and control groups. These reads have a median length of 22 bp and more than 82% map to miRbase. Preliminary analysis of retinal samples indicates a general decline in miRNA levels in affected individuals. Multiple miRNAs that determine translation of proteins involved in mitochondrial function, fibroblast response, and cell survival exhibited more than 2 fold decrease in GA patients compared to controls. This data will be validated by ongoing miRNA analysis of RPE samples and by complementary methods.

 
Conclusions:
 

Differential miRNA abundance measured in the retina and RPE of geographic atrophy patients may contribute to the development of age-related macular degeneration.

 
Keywords: age-related macular degeneration • genetics • gene/expression 
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