C3 Transferase-Expressing scAAV2 Transduces Ocular Anterior Segment Tissues and Lowers Intraocular Pressure in Mouse and Monkey

Abstract

Glaucoma is a lifelong disease with elevated intraocular pressure (IOP) as the main risk factor, and reduction of IOP remains the major treatment for this disease. However, current IOP-lowering therapies are far from being satisfactory. We have demonstrated that the lentivirus-mediated exoenzyme C3 transferase (C3) expression in rat and monkey eyes induced relatively long-term IOP reduction. We now show that intracameral injection of self-complementary AAV2 containing a C3 gene into mouse and monkey eyes resulted in morphological changes in trabecular meshwork and IOP reduction. The vector-transduced corneal endothelium and the C3 transgene expression, not vector itself, induced corneal edema as a result of actin-associated endothelial barrier disruption. There was a positive (quadratic) correlation between measured IOP and grade of corneal edema. This is the first report of using an AAV to transduce the trabecular meshwork of monkeys with a gene capable of altering cellular structure and physiology, indicating a potential gene therapy for glaucoma.

scAAV2-Mediated C3 Transferase Gene Therapy in a Rat Model with Retinal Ischemia/Reperfusion Injuries

“…5,40 Furthermore, as long as the serotype of AAV is considered, our previous studies demonstrated that intracameral injection of scAAV2-C3 induced a longterm IOP-lowering effect in monkey eyes. 41 Bogner et al 42 suggested that scAAV2-based vectors transduced the TM of mice and rats more effectively than did scAAV8-based vectors. In addition, the scAAV2 vector transduces RGCs around the optic nerve head more efficiently than do scAAV5 and scAAV8 after intravitreal injection.…”

“…Vector Construction scAAV2-EGFP (1 Â 10 12 viral genomes [vg]/mL) and scAAV2-C3 (5 Â 10 11 vg/mL) vectors were prepared by the Beijing FivePlus Molecular Medicine Institute (FMMI) (Beijing, China), as previously described. 41…”

RhoA/ROCK-YAP/TAZ Axis Regulates the Fibrotic Activity in Dexamethasone-Treated Human Trabecular Meshwork Cells

“…In addition, RhoA induced ECM accumulation and activated myofibroblast-like cells. These findings indicated that DEX may induce the fibrotic activity in HTM cells through the RhoA/ROCK pathway and that RhoA probably serves as an intervening factor for lowering IOP ( Rao et al, 2017 ; Tan et al, 2018 ; Tan et al, 2019 ; Chen et al, 2020 ; Tan et al, 2020 ).…”

“…RhoAG14V, a constitutively active variant of RhoA, has been reported to induce ECM synthesis ( Pattabiraman and Rao, 2010 ) and decrease AH outflow through the TM in anterior chambers ( Pattabiraman et al, 2015 ). Other studies have suggested that the RhoA/ROCK plays an important role in regulating IOP, which was reduced in the presence of RhoA inhibitor C3 transferase ( Tan et al, 2018 ; Tan et al, 2019 ; Tan et al, 2020 ) and ROCK inhibitor Y-27632 ( Pattabiraman et al, 2015 ; Chen et al, 2020 ). Based on these findings, it was demonstrated that DEX not only increased the expression level of RhoA but also activated ROCK.…”

“…Cell-matrix mechanoreceptors including integrin and cytoskeletal proteins play an important role in DEX-induced ECM remodeling ( Yemanyi et al, 2020 ). RhoA acting as one of the cytoskeletal proteins regulates the dynamic assembly of cytoskeleton through activating Rho-associated coiled-coil containing protein kinase (ROCK) ( Tan et al, 2018 ; Tan et al, 2019 ; Tan et al, 2020 ). ROCKs are the major downstream effectors of Rho GTPase and ROCK activation regulates the clustering of actin stress fiber in the cytoskeleton, which promotes actomyosin contractility and endogenous tension ( Rao and Epstein, 2007 ; Tian et al, 2009 ).…”