Topical delivery of a small molecule RUNX1 transcription factor inhibitor for the treatment of proliferative vitreoretinopathy

Santiago Delgado-Tirado,Heuy-Ching Wang,Eric Kim,John B Miller,Natalia Chmielewska,Elizabeth J Rossin,David Leyton-Cifuentes,Leo A Kim,Dean Eliott,Guannan Zhao,Daniela Isaacs-Bernal,Leslie Ramos,Dhanesh Amarnani,Lucía González-Buendía ,Whitney Greene ,Brandon V Duffy ,José M Ruiz-Moreno ,Said Arévalo-Alquichire ,Hannah Whitmore ,Joseph F Arboleda‐Velásquez

doi:10.1038/s41598-020-77254-0

Abstract

Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment surgery failure. Despite significant advances in vitreoretinal surgery, it still remains without an effective prophylactic or therapeutic medical treatment. After ocular injury or retinal detachment, misplaced retinal cells undergo epithelial to mesenchymal transition (EMT) to form contractile membranes within the eye. We identified Runt-related transcription factor 1 (RUNX1) as a gene highly expressed in surgically-removed human PVR specimens. RUNX1 upregulation was a hallmark of EMT in primary cultures derived from human PVR membranes (C-PVR). The inhibition of RUNX1 reduced proliferation of human C-PVR cells in vitro, and curbed growth of freshly isolated human PVR membranes in an explant assay. We formulated Ro5-3335, a lipophilic small molecule RUNX1 inhibitor, into a nanoemulsion that when administered topically curbed the progression of disease in a novel rabbit model of mild PVR developed using C-PVR cells. Mass spectrometry analysis detected 2.67 ng/mL of Ro5-3335 within the vitreous cavity after treatment. This work shows a critical role for RUNX1 in PVR and supports the feasibility of targeting RUNX1 within the eye for the treatment of an EMT-mediated condition using a topical ophthalmic agent.

Highlights

Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment surgery failure
We developed an ex-vivo model of PVR by growing fragments of freshly isolated human PVR membranes in Matrigel to examine a potential role for RUNX1 in growth of PVR membranes
We observed almost no outgrowths in samples treated with Ro5-3335 (150 μM), a small molecule RUNX1 inhibitor[54] (Fig. 1c), and the few outgrowths present were significantly shorter after 7 days (136.9 ± 18.9 μm and 180.3 ± 27.9 μm) (**p < 0.01)

Summary

Introduction

Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment surgery failure. After ocular injury or retinal detachment, misplaced retinal cells undergo epithelial to mesenchymal transition (EMT) to form contractile membranes within the eye. Proliferative vitreoretinopathy (PVR) is a condition in which, after retinal detachment or ocular trauma, some retinal cells are displaced from their anatomical location, undergo epithelial to mesenchymal transition (EMT), and grow uncontrollably beneath or on top of the retina triggering the formation of retinal membranes, tractional retinal detachment, and permanent vision loss[1,2]. Upon retinal detachment or trauma, RPE cells are misplaced from their anatomical location and induced to undergo EMT under the stimuli of growth factors, inflammatory cytokines, and exposure to vitreous, a collagenous gel that fills the space between the lens and the r etina[22,25].

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Results

Conclusion