September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
AMPK Activation for the Treatment of Fuchs Endothelial Corneal Dystrophy
Author Affiliations & Notes
  • Guadalupe Villarreal
    Ophthalmology, Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore, Maryland, United States
  • Tetsuya Toyono
    Ophthalmology, Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore, Maryland, United States
  • Laura KALLAY
    Ophthalmology, Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore, Maryland, United States
  • Albert S Jun
    Ophthalmology, Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore, Maryland, United States
  • Footnotes
    Commercial Relationships   Guadalupe Villarreal, Johns Hopkins University, P13615-01 (P); Tetsuya Toyono, None; Laura KALLAY, None; Albert Jun, Johns Hopkins University, P13615-01 (P)
  • Footnotes
    Support  Eye Bank Association of America/Richard Lindstrom Research Grant Award
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5283. doi:
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    • Get Citation

      Guadalupe Villarreal, Tetsuya Toyono, Laura KALLAY, Albert S Jun; AMPK Activation for the Treatment of Fuchs Endothelial Corneal Dystrophy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5283.

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

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Abstract

Purpose : Oxidant-antioxidant imbalance plays an important role in the pathogenesis of Fuchs endothelial corneal dystrophy (FECD). AMP-activated protein kinase (AMPK) has been shown to confer anti-oxidant properties in part via the upregulation of nuclear factor erythroid 2-related factor 2 (NRF2), a transcription factor critical for anti-oxidant cellular defense. The role of AMPK in the corneal endothelium remains unknown. We hypothesized that AMPK activation induces NRF2-dependent anti-oxidant transcriptional programs critical for the suppression of DNA damage, thereby establishing a corneal endothelial protective phenotype.

Methods : AMPK subunit expression in normal immortalized human corneal endothelial cells (HCECi) was examined by immunoblot analysis. HCECi were treated with varying doses, including 25, 100, 250, and 500 μM, of a small molecule, direct AMPK activator A-769662 or DMSO control for 24 hours. The effects of A-769662 in immortalized human Fuchs corneal endothelial cells (FECDi) were also investigated. Changes in mRNA levels of NRF2 as well as its target genes heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase quinone 1 (NQO1) were assessed by real-time quantitative PCR.

Results : AMPK expression was detected by immunoblot in HCECi. NRF2 mRNA levels were signficantly increased in HCECi by 16.4 ± 4.8% (n=3; p<0.05) and 94.4 ± 20.0% (n=3; p<0.01) compared to control cells following 24 hour incubation with 100 and 500 μM A-769662. Levels of HO-1 mRNA were significantly upregulated by 44.1 ± 8.0% (n=3; p<0.01), 111.4 ± 30.0% (n=3; p<0.05), and 349.0 ± 91.6% (n=3; p<0.05) compared to control cells following incubation with 100, 250, and 500 μM A-769662. Similarly, NQO1 mRNA levels were dose-dependently upregulated by 17.6 ± 1.7% (n=3; p<0.001), 72.1 ± 10.0% (n=3; p<0.01), and 185.8 ± 20.0% (n=3; p<0.001) with 100, 250, and 500 μM A-769662. While incubation of FECDi for 24 hours with 250 μM A-769662 did not result in a significant induction in NRF2 mRNA levels compared to control cells, HO-1 and NQO1 mRNA levels were significantly upregulated by 3,084.0 ± 278.0% (n=3; p<0.001) and 67.0 ± 22.5% (n=3; p<0.05).

Conclusions : Our data identify AMPK as an important regulator of anti-oxidant gene expression in normal and Fuchs human corneal endothelial cells, and suggest that pharmacological activation of AMPK may serve as a novel therapeutic approach for the establishment of a corneal endothelial protective phenotype.

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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