Innovative Gene Editing Approach for Glaucoma Treatment Home Innovative Gene Editing Approach For Glaucoma Treatment Researchers in the Zode lab are studying the use of a novel approach to treating glaucoma, an eye disease that can cause vision loss. Unfortunately, current treatments do not target the root cause of the problem. This research focuses on using base gene editing in cases caused by mutations in the myocilin (MYOC) gene, which is responsible for most occurrences of juvenile-onset glaucoma. This innovative method would remove the faulty gene that makes the harmful protein. Young children with a MYOC mutation experience symptoms of high intraocular pressure due to damage to eye cells in the trabecular meshwork tissue and progress rapidly toward vision loss. Knocking out MYOC selectively from the trabecular meshwork via genome editing provides an ideal strategy to provide a one-time “cure” for this disease. Current studies focus on precision genome editing that uses the traditional Cas9 system. A major bottleneck for genome-editing clinical applications for glaucoma is the lack of a robust and selective tissue-specific delivery system. Traditional CRISPR/Cas9-based gene knockouts introduce DNA double-strand breaks, which pose serious concerns. Base editors are more precise and have a lower risk of off-target effects. The research proposes using lipid nanoparticles to deliver adenine base editors to the trabecular meshwork tissue, aiming to knock out the mutant MYOC gene. Figure 1: Precision genome editing via base editors. CRISPR/Cas9 can be easily programmed to target any gene by the appropriate selection of guide RNAs. However, traditional CRISPR/Cas9-based gene knockouts introduce DNA double-strand breaks, which pose serious concerns. Recent advances in base editors allow us to change DNA precisely without any serious off-target effects. Figure 2: Lipid nanoparticles carrying mRNA expressing base editors and guide RNA. Figure 3: Lipid nanoparticles carrying Cre mRNA show robust activity in mouse trabecular meshwork. Arrows show trabecular meshwork. The Zode lab is well-versed in utilizing lipid nanoparticle-mediated mRNA delivery approaches for eye diseases. Lipid nanoparticles are designed to carry CRISPR-Cas9 to the trabecular meshwork and edit the targeted gene, minimizing inflammation and off-target effects (Figure 2). In addition, this system is very effective in delivering mRNA to the trabecular meshwork in living animals (Figure 2). Previous studies in the Zode lab have shown that lipid nanoparticles are highly efficient and selective in delivering cargo to trabecular meshwork cells (Figure 3). This delivery method could overcome current limitations in gene therapy for eye diseases, allowing for more targeted and effective treatments. The team is currently studying various lipid nanoparticles to deliver therapeutic ribonucleoprotein (RNP) base editors to trabecular meshwork cells. The major goals are 1) to validate the editing efficiency of these base editors in living mouse and human tissues; 2) to determine whether these base editors reduce MYOC in the trabecular meshwork and rescue glaucoma in a mouse model; 3) to establish the safety of these base editors in mice; and 4) to test these therapies in glaucoma patients. This approach will revolutionize glaucoma therapy by directly interfering with damage to the trabecular meshwork and provide a one-time therapy to treat glaucoma.