September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Reactive Oxygen Species (ROS) Production in ARPE-19 Cybrids Containing Mitochondria From AMD Versus Age-matched Non-AMD Controls Treated with Aflibercept, Ranibizumab and Bevacizumab
Author Affiliations & Notes
  • Rodrigo Costa
    Ophthalmology, UCI, IRVINE, California, United States
    ophthalmology, Instituto Donato Oftalmologia, Pocos de Caldas, MG, Brazil
  • Javier Cáceres-del-Carpio
    Ophthalmology, UCI, IRVINE, California, United States
  • Kunal Thaker
    Ophthalmology, UCI, IRVINE, California, United States
  • Tej Patel
    Ophthalmology, UCI, IRVINE, California, United States
  • Mohamed Abdelhamid Mohamed
    Ophthalmology, UCI, IRVINE, California, United States
  • Abdul Sami Memon
    Ophthalmology, UCI, IRVINE, California, United States
  • Jon Lucas Norman
    Ophthalmology, UCI, IRVINE, California, United States
  • M. Tarek Moustafa
    Ophthalmology, UCI, IRVINE, California, United States
  • Cristina M Kenney
    Ophthalmology, UCI, IRVINE, California, United States
  • Baruch D Kuppermann
    Ophthalmology, UCI, IRVINE, California, United States
  • Footnotes
    Commercial Relationships   Rodrigo Costa, None; Javier Cáceres-del-Carpio, None; Kunal Thaker, None; Tej Patel, None; Mohamed Mohamed, None; Abdul Memon, None; Jon Norman, None; M. Tarek Moustafa, None; Cristina Kenney, None; Baruch Kuppermann, AcuFocus (C), Alcon (C), Alimera (C), Allegro (C), Allergan (F), Ampio (C), Aqua Therapeutics (C), Bausch&Lomb (C), Genentech (F), Glaukos (C), Glaxo Smith Kline (F), Novagali (C), Novartis (C), Ophthotech (F), Regeneron (F), SecondSight (C), Staar (C), Surgical (C), Teva (C), Thrombogenics (F)
  • Footnotes
    Support  This work was supported by 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, David & Julianna Pyott Pan-American-Retina Research Fellowship.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5020. doi:
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    • Get Citation

      Rodrigo Costa, Javier Cáceres-del-Carpio, Kunal Thaker, Tej Patel, Mohamed Abdelhamid Mohamed, Abdul Sami Memon, Jon Lucas Norman, M. Tarek Moustafa, Cristina M Kenney, Baruch D Kuppermann; Reactive Oxygen Species (ROS) Production in ARPE-19 Cybrids Containing Mitochondria From AMD Versus Age-matched Non-AMD Controls Treated with Aflibercept, Ranibizumab and Bevacizumab. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5020.

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

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Abstract

Purpose : To determine if in vitro AMD (age-related macular degeneration) and non-AMD cybrids treated with different concentrations of anti-angiogenic drugs had similar levels of reactive oxygen species (ROS) production, a marker of cellular stress and injury.

Methods : Cybrids were created by fusing mitochondria-depleted ARPE-19 cells with platelets from subjects with AMD (n=5) or non-AMD controls (n=5). These cybrids have identical nuclei but different mitochondrial (mt) DNA. Cybrids were plated in 24-well plates, incubated for 48 hours, and then treated with aflibercept, ranibizumab or bevacizumab at 1X, 2X, 4X, or 10X clinical concentrations. After 24 hours, ROS production was measured with the H2DCF-DA assay. Results were normalized to untreated cells. All the experiments were repeated seven times.

Results : AMD and non-AMD cybrids treated with aflibercept did not show statistically significant changes at 1X and 2X concentrations, compared with untreated cells. ROS levels were decreased in the non-AMD group at 4X (P=0.013) and 10X (P=0.0002) concentrations and in the AMD group at 10X (P=0.0002) concentration. Cybrids treated with ranibizumab showed reduced ROS levels in the non-AMD group at 2X (P=0.0003), 4X (P=0.0007) and 10X (P=0.0004) concentrations. AMD cybrids showed a reduction in ROS levels at 4X (P=0.0007) and 10X (P=<0.0001) concentrations. After treatment with bevacizumab, non-AMD cybrids had reduced ROS levels at 1X (P=0.0037), 2X (P=0.0003), 4X (P=<0.0001) and 10X (P=<0.0001) concentrations, and the AMD cybrids showed a reduction in ROS levels at 2X (P=0.0018), 4X (P=<0.0001) and 10X (P=<0.0001) concentrations.

Conclusions : Cybrids containing identical nuclear genes but either AMD or non-AMD mitochondria, demonstrate differential responses to anti-VEGF drugs. Low concentrations of ranibizumab and bevacizumab had a protective effect in non-AMD cybrids, while the AMD cybrid group was not protected to the same extent. Further investigation is needed to better understand the interactions between anti-VEGF induced cellular protection and AMD mitochondria.

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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