Nucleoside Reverse Transcriptase Inhibitors Suppress Laser-Induced Choroidal Neovascularization in Mice

  • Invest Ophthalmol Vis Sci. 2015 Nov;56(12):7122-9. doi: 10.1167/iovs.15-17440.
Takeshi Mizutani  1 ,  Benjamin J Fowler  1 ,  Younghee Kim  1 ,  Reo Yasuma  1 ,  Laura A Krueger  1 ,  Bradley D Gelfand  2 ,  Jayakrishna Ambati  3
Affiliations
  • 1. Department of Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States.
  • 2. Department of Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States 2Department of Biomedical Engineering, University of Kentucky, Lexington, Kentucky, United States 3Department of Microbiology, Immunology, and Mole.
  • 3. Department of Ophthalmology and Visual Sciences, University of Kentucky, Lexington, Kentucky, United States 4Department of Physiology, University of Kentucky, Lexington, Kentucky, United States.
PMID: 26529046 DOI: 10.1167/iovs.15-17440
Abstract

Purpose: To evaluate the efficacy of nucleoside Reverse Transcriptase inhibitors (NRTIs) in the laser-induced mouse model of choroidal neovascularization (CNV).

Methods: We evaluated the NRTIs lamivudine (3TC), zidovudine (AZT), and abacavir (ABC) and the P2X7 antagonist A438079. Choroidal neovascularization was induced by laser injury in C57BL/6J wild-type, Nlrp3-/-, and P2rx7-/- mice, and CNV volume was measured after 7 days by confocal microscopy. Drugs were administered by intravitreous injection immediately after the laser injury. Vascular endothelial growth factor-A in RPE-choroid lysates was measured 3 days after laser injury by ELISA. HEK293 cells expressing human and mouse P2X7 were exposed to the selective P2X7 Receptor Agonist 2', 3'-(benzoyl-4-benzoyl)-ATP (Bz-ATP) with or without 3TC, and VEGF-A levels in media were measured by ELISA.

Results: Intravitreous injection of 3TC, AZT, and ABC significantly suppressed laser-induced CNV in C57BL/6J wild-type and Nlrp3-/- mice (P < 0.05) but not in P2rx7-/- mice. Intravitreous injection of A438079 also suppressed the laser-induced CNV (P < 0.05). The NRTIs 3TC, AZT, and ABC blocked VEGF-A levels in the RPE/choroid after laser injury in wild-type (P < 0.05) but not P2rx7-/- mice. Moreover, there was no additive effect of 3TC on CNV inhibition when coadministered with a neutralizing VEGF-A antibody. Stimulation of human and mouse P2X7-expressing HEK293 cells with Bz-ATP increased VEGF secretion (P < 0.001), which was abrogated by 3TC (P < 0.001). Stimulation of primary human RPE cells with Bz-ATP increased VEGFA and IL6 mRNA levels, which were abrogated by 3TC.

Conclusions: Multiple clinically relevant NRTIs suppressed laser-induced CNV and downregulated VEGF-A, via P2X7.

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