Neuroprotective miRNA delivery via extracellular vesicles to glaucomatous retina

Durmaz, Esmahan 2024. Neuroprotective miRNA delivery via extracellular vesicles to glaucomatous retina. PhD Thesis, Cardiff University. Item availability restricted.

	PDF (Thesis) - Accepted Post-Print Version Restricted to Repository staff only until 12 February 2026 due to copyright restrictions. Download (8MB)
	PDF (Cardiff University Electronic Publication Form) Restricted to Repository staff only Download (130kB)

Abstract

Retinal diseases are the leading cause of blindness, resulting in irreversible degeneration and death of retinal neurons. One such cell type, the retinal ganglion cell (RGC), is responsible for connecting the retina to the rest of the brain through its axons that make up the optic nerve and is the primary cell lost in glaucoma and traumatic optic neuropathy. To date, different therapeutic strategies have been investigated to protect RGCs from death and preserve vision, yet current strategies are restricted to treating neuron loss by targeting the raised intraocular pressure in glaucoma. Therefore, the overall aim of the thesis was to explore novel therapeutic strategies for RGC degeneration caused by glaucoma, the background of which is explored in Chapter 1. Chapter 2 tested 4 different miRNA-based treatment strategies for RGC degeneration, delivered through a commercial liposome-based vehicle. The results showed that miRNA 181a-3p provided neuroprotection and higher Ca2+ activity. In silico investigation of possible mRNAs modulated by these miRNAs was performed. The effect of miRNA 181a-3p and miRNA 23a-3p on gene expression was investigated with qPCR and did not show a significant change in selected genes, which includes p38, IL1β, Bcl, Cas3, and FGF for miRNA 181a-3p and Wnt5a, calmodulin, calcineurin, CaMKII, NFAT, and Cas8 for miRNA-23a-3p. Chapter 3 investigated EV based therapeutics to mitigate RGC degeneration following IOP elevation. According to our results, R-28 (retinal progenitor cell line) derived EVs improved RGC survival in rat primary retinal cell cultures and human embryonic stem cell derived RGCs with RGC degeneration model induced by colchicine. Moreover, it provided RGC protection in an in vivo microbead glaucoma model. Alterations in miRNA content were also observed in RGCs after EV treatment. Chapter 4 aimed to bring together the findings of chapters 2 and 3, to develop an EV-based drug delivery system for miRNA therapeutic delivery to retinal cells. R-28 derived EVs (from Chapter 2) were successfully loaded with miRNAs (from Chapter 3) and delivered their cargo efficiently into retinal cells. The possible effect of electroporation was investigated on EV size, concentration, and charge, which showed that there was no significant difference between EVs before and after electroporation in terms of size and charge. However, the EV particle concentration was observed lower in the electroporated EV solution. This study investigated EV mediated RNA kinetics in the vitreous and showed a gradual decrease via EV delivery in vivo. These studies provide important information about the potential of miRNA therapeutics and R-28 derived EVs as a therapeutic and drug delivery system for retinal diseases. Consequently, miRNA 181a-3p might prevent RGCs from degeneration and R-28 derived EVs have therapeutic potential for RGC degeneration in glaucoma as well as represent an ideal drug delivery system for the retina.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Optometry and Vision Sciences
Funders:	National Ministry of Education, Turkey, Hodge ECR Grant, EGAS Fund
Date of First Compliant Deposit:	12 February 2025
Last Modified:	12 Feb 2025 15:50
URI:	https://orca.cardiff.ac.uk/id/eprint/176118

Actions (repository staff only)

Edit Item