June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
ROCK Inhibitors AR-13324 and AR-13154(S) Block Choroidal Angiogenesis and Protect the Barrier Function of Retinal Pigment Epithelium
Author Affiliations & Notes
  • Jindong Ding
    Aerie Pharmaceuticals, Inc., Durham, North Carolina, United States
  • Kevin Carbajal
    Aerie Pharmaceuticals, Inc., Durham, North Carolina, United States
  • Casey Kopczynski
    Aerie Pharmaceuticals, Inc., Durham, North Carolina, United States
  • Cheng-Wen Lin
    Aerie Pharmaceuticals, Inc., Durham, North Carolina, United States
  • Footnotes
    Commercial Relationships   Jindong Ding, Aerie Pharmaceuticals, Inc. (E); Kevin Carbajal, Aerie Pharmaceuticals, Inc. (E); Casey Kopczynski, Aerie Pharmaceuticals, Inc. (E); Cheng-Wen Lin, Aerie Pharmaceuticals, Inc. (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1967. doi:
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      Jindong Ding, Kevin Carbajal, Casey Kopczynski, Cheng-Wen Lin; ROCK Inhibitors AR-13324 and AR-13154(S) Block Choroidal Angiogenesis and Protect the Barrier Function of Retinal Pigment Epithelium
      . Invest. Ophthalmol. Vis. Sci. 2017;58(8):1967.

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

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Abstract

Purpose : A significant portion of patients with exudative age-related macular degeneration (AMD) does not respond to anti-vascular endothelial growth factor (VEGF) therapy, and many who do experience an improvement in vision eventually lose the benefit over time. It has been shown in preclinical models of exudative AMD that Rho-associated protein kinase (ROCK) inhibitors are effective as monotherapy and in combination with anti-VEGF agents in reducing choroidal neovascularization and vascular leakage. In present study, we tested the effects of AR-13324, a ROCK and norepinephrine transporter (NET) inhibitor, and AR-13154(S), a multi-kinase inhibitor of ROCK/PKC/JAK/PDGFR-b, on ex vivo choroidal angiogenesis and in vitro retinal pigment epithelium (RPE) barrier function.

Methods : Organotypic choroidal tissue was isolated from C57BL/6 mice and cultured in the Matrigel with addition of endothelium growth medium. Tissues were then treated with various concentrations of AR-13324 or AR-13154(S). Choroidal angiogenesis was quantified by measuring the area of the choroidal sprouting. In the RPE barrier function assay, primary porcine RPEs were cultured on transwell inserts and treated with 150μM of hydroquinone with or without various concentrations of AR-13324 and AR-13154(S). Transepithelial resistance (TER) was measured as the indicator of RPE barrier function.

Results : Choroidal vessels sprouted in the Matrigel in the presence of endothelium growth medium. The sprouting area was reduced dose-dependently in the presence of AR-13324 and AR-13154(S), each with an IC50 of approximately 1μM. Primary porcine RPEs established high TER (~ 800Ohm.cm2) after 2-weeks of culture in transwell inserts with polarization-inducing medium. Treatment of the RPEs with 10μM of AR-13324 or AR-13154(S) increased the TER by 60% without signs of cytotoxicity. Treatment with 150μM hydroquinone, a toxin found in cigarette smoke, decreased the TER by about 50%. This TER-lowering effect of hydroquinone was attenuated by both AR-13324 and AR-13154(S).

Conclusions : In ex vivo and in vitro assays, AR-13324 and AR-13154(S) inhibited choroidal angiogenesis and protected RPE barrier function. This study demonstrated that ROCK inhibitors are potential candidates for treatment of AMD.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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