Delivery of Antisense Oligonucleotides to the Mouse Retina.

Alejandro Garanto

doi:10.1007/978-1-0716-2010-6_22

Abstract

The eye is the organ in charge of vision and, given its properties, has become an excellent organ to test genetic therapies, including antisense oligonucleotide (AON) technology. In fact, the first AON receiving FDA and EMA approval was meant to treat an eye condition. Currently, dozens of clinical trials are being conducted for a variety of subtypes of inherited retinal disease. Although most of them are based on gene augmentation therapies, a phase 3 and two phase 1/2 clinical trials using AONs are ongoing. Since the retina is a layered structure of nondividing cells, obtaining human retinal tissue and expanding it in the lab is not possible, unless induced pluripotent stem cell technology is used. Mouse models have helped to elucidate the function of many genes, and the retinal structure is quite similar to that of humans. Thus, drug delivery to the mouse eye can provide valuable information for further optimization of therapies. In this chapter, the protocol for intravitreal injections of AONs is described in detail.

Highlights

Inherited retinal diseases (IRDs) are highly heterogeneous, but in general, the first symptoms appear during the first decade of life, and the disease progresses with age, often leading to total blindness
An example is Luxturna, an adeno-associated virus (AAV)-based therapy, which has become the first gene therapy treatment with market approval for an eye disease [3]. This achievement has sped up the development of many other potential treatments currently under investigation at both preclinical and clinical levels for a variety of retinal diseases
We developed a humanized mouse model for the intronic variant c.2991+1655A>G in CEP290 [6]

Summary

Introduction

The eye is the window to the world around us. Vision occurs thanks to a thin neuronal layer located at the back of the eye called the retina. An example is Luxturna, an adeno-associated virus (AAV)-based therapy, which has become the first gene therapy treatment with market approval for an eye disease [3]. This achievement has sped up the development of many other potential treatments currently under investigation at both preclinical and clinical levels for a variety of retinal diseases. Another example of molecules that have shown promising results in humans is antisense oligonucleotides (AONs).

For Injections

For Harvesting Eyes

Preparation for the Procedure

The Procedure

After the Procedure