Preclinical Dose Escalation Study of Intravitreal AAV RS1 Gene Therapy in a Mouse Model of X linked Retinoschisis: Dose Dependent Expression and Improved Retinal Structure and Function.
Gene therapy for inherited retinal diseases has been shown to ameliorate functional and structural defects in both animal models and in human clinical trials. X linked retinoschisis (XLRS) is an early age onset macular dystrophy resulting from loss of an extracellular matrix protein (RS1) In preparation for a human clinical gene therapy trial, we conducted a dose range efficacy study of the clinical vector, a self complementary AAV delivering a human retinoschisin (RS1) gene under control of the RS1 promoter and an interphotoreceptor binding protein enhancer (AAV8 scRS/IRBPhRS) in the retinoschisin knockout (Rs1 KO) mouse. The therapeutic vector at 1 x 10(6) to 2.5 x 10(9) (1E6 2.5E9) vector genomes (vg)/eye or vehicle was administered to one eye of 229 male Rs1 KO mice by intravitreal injection at 22 3 days postnatal age (PN) Analysis of retinal function (dark adapted electroretinogram, ERG) structure (cavities and outer nuclear layer thickness) by in vivo retinal imaging using optical coherence tomography, and retinal immunohistochemistry (IHC) for RS1 was done 3 4 months and/or 6 9 months postinjection (PI) RS1 IHC staining was dose dependent across doses =1E7 vg/eye, and the threshold for significant improvement in all measures of retinal structure and function was 1E8 vg/eye. Higher doses, however, did not produce additional improvement. At all doses showing efficacy, RS1 staining in Rs1 KO mouse was less than that in wild type mice. Improvement in the ERG and RS1 staining was unchanged or greater at 6 9 months than at 3 4 months PI. This study demonstrates that vitreal administration of AAV8 scRS/IRBPhRS produces significant improvement in retinal structure and function in the mouse model of XLRS over a vector dose range that can be extended to a human trial. It indicates that a fully normal level of RS1 expression is not necessary for a therapeutic effect.
Human gene therapy