April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Histone Deacetylase Expression and Inhibition in Age Related Macular Degeneration
Author Affiliations & Notes
  • Mark Ellsworth Kleinman
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Andre Berner
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Kablian Mohan
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Ding-Yuan Lou
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Jennifer Brown
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Justin West
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Rei Kono
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
  • Ilene Sugino
    Institute of Ophthalmology & Visual Science, New Jersey Medical School, Newark, NJ
  • Marco Zarbin
    Institute of Ophthalmology & Visual Science, New Jersey Medical School, Newark, NJ
  • Jayakrishna Ambati
    Ophthalmology & Visual Sci, Univ of Kentucky, Lexington, KY
    Physiology, University of Kentucky, Lexington, KY
  • Footnotes
    Commercial Relationships Mark Kleinman, None; Andre Berner, None; Kablian Mohan, None; Ding-Yuan Lou, None; Jennifer Brown, None; Justin West, None; Rei Kono, None; Ilene Sugino, None; Marco Zarbin, None; Jayakrishna Ambati, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3457. doi:https://doi.org/
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      Mark Ellsworth Kleinman, Andre Berner, Kablian Mohan, Ding-Yuan Lou, Jennifer Brown, Justin West, Rei Kono, Ilene Sugino, Marco Zarbin, Jayakrishna Ambati; Histone Deacetylase Expression and Inhibition in Age Related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3457. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: The role of histone deacetylases (HDACs) in the postnatal control of cell survival and gene expression remains unclear. Reports of improved retinal function in retinitis pigmentosa and age-related macular degeneration (AMD) led us to investigate the effects of specific HDAC inhibition on the retinal pigment epithelium (RPE) in vitro and in vivo and analyze HDAC expression in situ.

Methods: Eyes with dry AMD and controls (n=3) were examined by immunohistochemistry for HDAC expression. Total RNA was harvested from RPE/choroid (n=3) and interrogated with Nanostring™ and real-time PCR. Wild-type (Wt) C57BL/6J mouse eyes (n=3) were evaluated with HDAC immunofluorescence. Mice (n=6) were imaged by fundus photography 1 week after intravitreous injection of HDAC inhibitors (suberoylanilide hydroxamic acid (SAHA), trichostatin-A (TSA)) and vehicle (DMSO). ZO-1 immuno-stained RPE/choroid flatmounts (n=3) were analyzed by fluorescent microscopy. RPE/choroid tissue (n=3) was harvested at 48 hours and analyzed by targeted PCR arrays (Taqman®). Primary human RPE (hRPE) isolates (n=6, individual donors) were treated with various HDAC inhibitors. Short-interfering RNAs targeting HDACs were also evaluated in vitro. Cell viability assays and targeted PCR arrays were performed. Statistical analyses were performed with non-parametric tests (Mann-Whitney U) with significance determined at P<0.05.

Results: HDAC-2 was down-regulated in eyes with AMD compared to controls (-1.45±0.23, P<0.05) and expressed in mouse RPE and in hRPE. Intravitreous injection of Class I/II HDAC inhibitors led to focal atrophy of RPE as observed on fundus imaging and ZO-1 immunofluorescence. HDAC inhibition in hRPE resulted in upregulated cell death and wide-spread changes in the cytokine expression profile. Similar disruptions in pro-inflammatory gene expression were observed in the mouse model and with specific targeting of HDAC-2 via RNA interference in vitro.

Conclusions: We found that Class I/II HDAC inhibition resulted in increased RPE cell death and inflammatory cytokine expression. Down-regulation of HDAC-2 was observed in RPE from eyes with AMD suggesting a role for this important epigenetic mechanism in the pathogenesis of this disease.

Keywords: 412 age-related macular degeneration • 533 gene/expression • 701 retinal pigment epithelium  
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