September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
HDAC1/2 are critical for RPE homeostasis and health
Author Affiliations & Notes
  • Mark Ellsworth Kleinman
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Kyung Jung
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Jacob Roney
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Subhash C Prajapati
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Dingyuan Lou
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Sushil Dubey
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Jennifer Brown
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Kabhilan Mohan
    Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States
  • Footnotes
    Commercial Relationships   Mark Kleinman, University of Kentucky (P), Vergent Biotech (C); Kyung Jung, None; Jacob Roney, None; Subhash Prajapati, None; Dingyuan Lou, None; Sushil Dubey, None; Jennifer Brown, None; Kabhilan Mohan, None
  • Footnotes
    Support  NIH K08EY021757, Foundation Fighting Blindness, and Research to Prevent Blindness
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 3155. doi:
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    • Get Citation

      Mark Ellsworth Kleinman, Kyung Jung, Jacob Roney, Subhash C Prajapati, Dingyuan Lou, Sushil Dubey, Jennifer Brown, Kabhilan Mohan; HDAC1/2 are critical for RPE homeostasis and health. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3155.

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

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Abstract

Purpose : Histone deacetylases are a family of 18 known members, classified in four groups based on their homology to yeast proteins. In previous RNA-seq data, we found decreased expression of HDAC1/2 in RPE/choroid samples from human eyes with advanced dry age-related macular degeneration (AMD). Here, we report on the development of experimental models of HDAC1/2 deficiency and discover a critical role for these epigenetic regulators in the homeostasis and health of the RPE.

Methods : Hdac1f/f, Hdac2f/f, and Hdac1/2f/f (gift of Eric Olson) mice were crossed with Vmd2-Cre (Jackson Labs). Conditional ablation of Hdac1/2 in RPE and localized Cre expression were confirmed by qPCR and immunofluorescence (IF). Fundus examination by multimodal imaging and ERG were performed (n=12-16). RPE morphology was examined by ZO-1 IF on RPE/choroidal flatmounts, and sections were analyzed by H&E staining (n=4-6). AAV2-iCre or control AAV2-Gfp sub-retinal injections were performed in WtC57BL/6J, Hdac1f/f, Hdac2f/f, and Hdac1/2f/f (n=4-6) followed by fundus examination at 14 days. Intravitreous injections of small molecule class I HDAC inhibitors and siRNAs were studied in WtC57BL/6J mice (n=6-8).

Results : Hdac1/2f/fx Vmd2Cre exhibited a robust phenotype characterized by pan-retinal degeneration with complete atrophy of the RPE by 1 month of age (Figure 1) and focal areas of depigmentation in their fur coat. Viable RPE was present at 2 weeks of age as shown in fundus imaging and ZO-1 IF. Loss of Hdac1/2 expression and Cre expression were confirmed in the RPE by qPCR and IF. At 1 month, SD-OCT revealed significant disruptions in the outer retina, and full-field ERGs had markedly diminished a- and b-wave amplitudes. Hdac1f/f and Hdac2f/f mice with Vmd2Cre demonstrated a less severe phenotype with focal RPE degeneration which developed at 2 months. AAV2-iCre (Figure 1) and siRNAs or small molecule inhibitors targeting HDAC1/2 induced focal areas of RPE degeneration as compared to control treatments.

Conclusions : Conditional ablation of Hdac1/2 in the mouse RPE resulted in severe degeneration and complete atrophy by 1 month. Similar results were achieved with postnatal ablation or inhibition of Hdac1/2 with siRNAs or small molecules. This pathway may be important in the pathogenesis of dry AMD, as the expression of Hdac1/2 is significantly decreased in the advanced stages of this disease.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

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