Development of mRNA-based therapeutics for proliferative vitreoretinopathy

Abstract

Purpose : The use of messenger RNA (mRNA)-based therapeutics in diseases involving transient expression of proteins is a rapidly expanding area. However, the use of this technology in ocular pathologies has yet been explored. Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment surgery failure which is characterized by the formation of contractile membranes that often require multiple surgeries and have no approved pharmacological treatments. Epithelial to mesenchymal transition (EMT) is a known driver of PVR progression therefore we screened anti-EMT targets using in vitro and in vivo models of PVR.

Methods : mRNA candidate therapeutics targeting specific factors associated with EMT for preventing fibrotic membrane formation in PVR were developed. The effects on cellular proliferation of these candidates were first tested in vitro, in patient-derived cultures from dissociated PVR membranes. Their ability to prevent fibrotic membrane formation was then evaluated in an in vivo rabbit model of PVR. Single cell RNA sequencing was carried out on eight, freshly dissociated PVR membranes.

Results : RUNX1, which is strongly expressed in PVR membrane cells was inhibited via expression of a dominant negative protein, RUNX1-Trap. Membrane proliferation was inhibited by 56% (P<0.05), with no associated cytotoxicity (P>0.05). The PVR severity score was significantly reduced in the rabbit model of PVR (control: 6.9±0.67 S.E.M vs RUNX1-Trap 3.67±1.1 S.E.M) (P<0.05), with reduced membrane formation and extracellular matrix deposition observed (Figure 1).

Conclusions : These data show preclinical therapeutic efficiency of mRNA-encoded dominant negative RUNX1-Trap in vivo for the first time and demonstrate the potential utility of mRNA-based therapeutics for ocular conditions. The results suggest that mRNA-encoded RUNX1-Trap may be a potential therapeutic candidate for ocular application in the treatment of PVR.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.

 

Figure 1: A) Single cell RNA sequencing of PVR membranes from eight patients. B) Rabbit model of PVR to test mRNA-based therapeutics. C) Hematoxylin and Eosin-stained samples showing reduced cell PVR membrane formation in the RUNX1-Trap treated rabbit eyes compared with control.