September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Effects of anti-angiogenic drugs on expression patterns of epigenetic acetylation pathway genes
Author Affiliations & Notes
  • Mohamed Mohamed
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
    Ophthalmology Department, Minia University, Minia, Egypt
  • Jon Lucas Norman
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Javier Cáceres-del-Carpio
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Rodrigo Costa
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Abdul Sami Memon
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • M. Tarek Moustafa
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
    Ophthalmology Department, Minia University, Minia, Egypt
  • Shari R Atilano
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Marilyn Chwa
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Cristina M Kenney
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Baruch D Kuppermann
    Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Mohamed Mohamed, None; Jon Norman, None; Javier Cáceres-del-Carpio, None; Rodrigo Costa, None; Abdul Memon, None; M. Tarek Moustafa, None; Shari Atilano, None; Marilyn Chwa, None; Cristina Kenney, None; Baruch Kuppermann, AcuFocus (C), Alcon (C), Alimera (C), Allegro (C), Allergan (C), Allergan (F), Ampio (C), Aqua Therapeutics (C), Bausch & Lomb (C), Genentech (C), Genentech (F), Glaukos (C), Neurotech (C), Novagali (C), Novartis (C), Ophthotech (C), Ophthotech (F), Regeneron (C), Regeneron (F), Thrombogenics (F)
  • Footnotes
    Support  Discovery Eye Foundation, Guenther Foundation, Beckman Initiative for Macular Research, Polly and Michael Smith Foundation, Max Factor Family Foundation, Iris and B. Gerald Cantor Foundation.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5028. doi:
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    • Get Citation

      Mohamed Mohamed, Jon Lucas Norman, Javier Cáceres-del-Carpio, Rodrigo Costa, Abdul Sami Memon, M. Tarek Moustafa, Shari R Atilano, Marilyn Chwa, Cristina M Kenney, Baruch D Kuppermann; Effects of anti-angiogenic drugs on expression patterns of epigenetic acetylation pathway genes. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5028.

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

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Abstract

Purpose : Epigenetic changes can affect cellular transcriptional activities and have been associated with retinal diseases. This study examines the effects of anti-angiogenic drugs on the transcription of acetylation genes in immortalized human RPE cells (ARPE-19) in vitro, to determine if epigenetic pathways are modulated by anti-angiogenic treatment.

Methods : ARPE-19 cells were treated with ranibizumab, bevacizumab, or aflibercept in 1X and 2X concentrations of the clinical intravitreal dose. Untreated cells were used as a control. RNA was isolated and reverse transcribed into copy DNA (cDNA). Quantitative polymerase chain reaction (Q-PCR) was performed in triplicates using primers for acetylation genes: HAT1 (histone acetyltransferase 1), and HDACs (histone deacetylases) 1, 6, and 11. HPRT1 (hypoxanthine phosphorbosyltransferase 1) and HMBS (hydroxymethylbilane synthase) were used as housekeeping genes. ΔΔCt (differences in cycle thresholds) was obtained and folds calculated using the formula 2ΔΔCt. Unpaired t test was used for statistical analysis.

Results : Bevacizumab-treated cells expressed 0.73-fold of HDAC1 (p=0.0001), 1.3-fold of HDAC6 (p=0.004), and 1.52-fold of HDAC11 (p=0.009) at 1X concentration, and 1.39-fold of HDAC11 (p=0.013) at 2X concentration. Aflibercept-treated cells showed 0.74-fold of HDAC1 (p=0.0002), 1.11-fold of HDAC6 (p=0.037), and 1.27-fold of HDAC11 (p=0.02) at 1X concentration, and 1.48-fold of HDAC11 (p=0.023) at 2X concentration. Ranibizumab-treated cells expressed 1.26-fold of HDAC6 (p=0.018) at 1X concentration, and HDAC11 (1.32-fold, p=0.006) at 2X concentration. Untreated samples were assigned a value of 1. No statistically significant difference in HAT1 expression was found in any group.

Conclusions : Our results suggest that anti-angiogenic drugs can alter expression levels of acetylation genes in ARPE-19 cells. This is significant because the degree of histone acetylation regulates chromatin structure and function, which can impact transcriptional activities for numerous genes involved in age-related macular degeneration (AMD) pathogenesis. Our findings also suggest anti-VEGF drugs may have broader mechanisms of action than just binding vascular endothelial growth factor (VEGF) and blocking receptor interactions for angiogenesis inhibition.

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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