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Siddharth Patel, Renee Christine Ryals, Mark E Pennesi, Gaurav Sahay; Messenger RNA Therapeutics for Gene Delivery in the Retina. Invest. Ophthalmol. Vis. Sci. 2018;59(9):5333.
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© ARVO (1962-2015); The Authors (2016-present)
The advent of messenger RNA (mRNA) therapeutics has accelerated gene therapy research. mRNA therapeutics can treat myriad diseases which were once considered undruggable. Advances in mRNA technology have unlocked high efficacy gene delivery in hard-to-transfect retinal cells for intracellular delivery of genes, gene silencing, or genome editing. Moreover, mRNA poses no risk of genomic integration and has been shown to have a higher efficiency at gene delivery relative to DNA by avoiding the necessity for nuclear entry. Additionally, it provides rapid and transient protein expression. Lipid-based nanoparticles (LNPs) are capable of encapsulating and delivering mRNA intracellularly, are easy to scale up, and have shown excellent clinical efficacy and safety. Nanoparticle-based mRNA delivery provides an excellent opportunity for therapeutic intervention of the numerous genetic disorders affecting the different layers of the retina.
In this study, 1) we designed LNPs that encapsulated negatively charged mRNA and showed high efficacy in vivo transfection of the RPE cells. 2) We are screening bioactive lipids to guide the nanoparticles towards different cells in the retinal layer.
We have developed LNPs that can successfully deliver reporter gene, luciferase and EGFP (Fig. 1), mRNA to the RPE cells in mice following intravitreal injections. Additionally, we have discovered bioactive lipids that can enhance gene delivery in vitro and in vivo. Further studies are ongoing to target different cells in the retina using these bioactive lipids as constituents of our LNPs.
We have developed lipid-based nanoparticles with the capability to deliver mRNA for gene delivery to the retina. mRNA showed potent gene expression within 4 hours and peaked at 24 hours. Engineering of different LNPs can prove effective in the cell-specific intracellular delivery of genes in multiple layers of the retina.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
Figure 1: (A-B) Fundus images showing retina transfected with LNP-mediated EGFP mRNA and control eye injected with PBS. (C-D) Immunohistochemistry of the transfected retina demonstrating EGFP expression localized to the RPE cells.
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