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Abdul Sami Memon, Javier Cáceres-del-Carpio, Rodrigo Costa, Mohamed Mohamed, M. Tarek Moustafa, Kunal Thaker, Theresa Thai, Sonali R Nashine, Marilyn Chwa, Cristina M Kenney, Baruch D Kuppermann; Response of ARPE-19 cybrids of European and African origin to high glucose and bevacizumab (Avastin). Invest. Ophthalmol. Vis. Sci. 2016;57(12):5057.
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© ARVO (1962-2015); The Authors (2016-present)
Individuals of African-origin are more susceptible to diabetes mellitus and its complications than those of European-origin. Worldwide, bevacizumab is the most commonly used drug for diabetic macular edema and proliferative diabetic retinopathy. The purpose of this study was to determine if cybrids, cultured cells created with identical nuclei but mitochondrial DNA (mtDNA) from either European-origin (H haplogroup) or African-origin (L haplogroup) subjects, showed differences in reactive oxygen species (ROS) production, marker for oxidative stress, after exposure to elevated glucose concentration ± bevacizumab (Avastin).
H and L cybrids were created by fusing immortalized human retinal pigment epithelial (ARPE-19) cells devoid of mtDNA with platelets isolated from subjects with either H or L haplogroups mtDNA. Cybrids were cultured for 24 hours in low glucose media (LGM, 17mM) or high glucose media (HGM, 35mM) with or without bevacizumab at 1x clinical dose, equivalent to 0.05ml containing 1.25mg of bevacizumab injected in 4ml of vitreous. ROS production was measured by the H2DCF-DA assay. Data were analyzed using GraphPad Prism version 5.01.
When exposed to bevacizumab, H cybrids in LGM and HGM had significantly decreased ROS levels (76.6%±5.1, P=0.0007 and 68.8%±4.6, P=<0.0001, respectively) compared to LGM cultures (100%±2.9), while no significant change was observed in HGM-treated cultures.L cybrids cultured in HGM, LGM+bevacizumab and HGM+bevacizumab had significantly decreased ROS levels (78.8%±1.8, P=<0.0001, 81.7%±3.5, P=0.0001 and 66.6%±3.4, P<0.0001, respectively) compared to LGM cultures (100%±2.4).
Our results showed the ROS levels declined in H and L cybrids after exposure to bevacizumab in both LGM and HGM-treated cultures and L cybrids also showed decreased ROS level in HGM-treated cultures, suggesting that this drug has a protective effect in these cells.
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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