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Javier Cáceres-del-Carpio, Shari R Atilano, Jon Lucas Norman, M. Tarek Moustafa, Rodrigo Costa, Abdul Sami Memon, Mohamed Abdelhamid Mohamed, G. Astrid Limb, Cristina M Kenney, Baruch D Kuppermann; Anti-VEGF Drugs can Modify Gene Expression in Retinal Müller Cells in Vitro. Invest. Ophthalmol. Vis. Sci. 2016;57(12):772.
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© ARVO (1962-2015); The Authors (2016-present)
Anti-vascular endothelial growth factor (VEGF) therapy is the most commonly used treatment for neovascular age-related macular degeneration, diabetic macular edema and macular edema due to retinal vein occlusion. Ranibizumab and aflibercept are approved by the Food and Drug Administration for those diseases; bevacizumab is used off-label worldwide, and there are some reports of ziv-aflibercept use in small series of patients. The purpose of this study is to assess the in vitro differences in gene expression of several key anti-oxidant, pro apoptotic and angiogenesis-related genes in cultured human retinal Müller cells (MIO-M1) treated with different concentrations of anti-VEGF agents.
MIO-M1 cells were plated in 6-well plates and then treated with 1x and 2x clinical dose of ranibizumab, bevacizumab, aflibercept and ziv-aflibercept for 24 hours. RNA was isolated, cDNA synthesized and quantitative polymerase chain reaction (Q-PCR) performed using primers for anti-oxidant genes: glutathione peroxidase 3 (GPX3) and superoxide dismutase 2 (SOD2); pro apoptotic genes: B-cell lymphoma 2 like 13 (BCL2L13) and BCL2-associated X protein (BAX); and angiogenesis-related genes: vascular endothelial growth factor A (VEGFA) and placental growth factor (PGF or PlGF). ΔΔCT (Differences in cycle threshold) values were calculated, and folds were obtained with the formula 2ΔΔCt.
Significant upregulation for GPX3 gene was found in 1x and 2x doses of ranibizumab, bevacizumab and aflibercept. Significant downregulation of SOD2 gene was found in both concentrations in all drugs studied. BCL213 gene was found significantly upregulated in ranibizumab 1x, bevacizumab 1x/2x, aflibercept 1x/2x, and ziv-aflibercept 1x. No significant differences for BAX gene were found except for bevacizumab 1x and ziv-aflibercept 1x/2x. VEGFA was found significantly down regulated in both concentrations of all the drugs studied except in ziv-aflibercept 2x. PlGF was found significantly upregulated in 1x/2x bevacizumab and aflibercept, and significantly downregulated in ziv-aflibercept 1x/2x. (Table 1)
Anti-VEGF drugs in MIO-M1 cells can change the gene expression of pathways distinct from VEGF, including anti-oxidant and pro-apoptotic genes. Bevacizumab as well as aflibercept can upregulate PlGF. Ziv-aflibercept appears to be less efficient in regulating VEGF and PlGF.
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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