Formulation and efficacy of ECO/pRHO-ABCA4-SV40 nanoparticles for nonviral gene therapy of Stargardt disease in a mouse model.

Da Sun, Wenyu Sun, Song-Qi Gao, Cheng Wei, Amirreza Naderi, Andrew L Schilb, Josef Scheidt, Sangjoon Lee, Timothy S Kern, Krzysztof Palczewski, Zheng-Rong Lu

Journal of controlled release : official journal of the Controlled Release Society 2021 Feb 10

filter terms:

It is still a challenge to develop gene replacement therapy for retinal disorders caused by mutations in large genes, such as Stargardt disease (STGD). STGD is caused by mutations in ABCA4 gene. Previously, we have developed an effective non-viral gene therapy using self-assembled nanoparticles of a multifunctional pH-sensitive amino lipid ECO and a therapeutic ABCA4 plasmid containing rhodopsin promoter (pRHO-ABCA4). In this study, we modified the ABCA4 plasmid with simian virus 40 enhancer (SV40, pRHO-ABCA4-SV40) for enhanced gene expression. We also prepared and assessed the formulations of ECO/pDNA nanoparticles using sucrose or sorbitol as a stablilizer to develop consistent and stable formulations. Results demonstrated that ECO formed stable nanoparticles with pRHO-ABCA4-SV40 in the presence of sucrose, but not with sorbitol. The transfection efficiency in vitro increased significantly after introduction of SV40 enhancer for plasmid pCMV-ABCA4-SV40 with a CMV promoter. Sucrose didn't affect the transfection efficiency, while sorbitol resulted in a fluctuation of the in vitro transfection efficiency. Subretinal gene therapy in Abca4-/- mice using ECO/pRHO-ABCA4 and ECO/pRHO-ABCA4-SV40 nanoparticles induced 36% and 29% reduction in A2E accumulation respectively. Therefore, the ECO/pABCA4 based nanoparticles are promising for non-viral gene therapy for Stargardt disease and can be expended for applications in a variety of visual dystrophies with mutated large genes. Copyright © 2020 Elsevier B.V. All rights reserved.

Citation

Da Sun, Wenyu Sun, Song-Qi Gao, Cheng Wei, Amirreza Naderi, Andrew L Schilb, Josef Scheidt, Sangjoon Lee, Timothy S Kern, Krzysztof Palczewski, Zheng-Rong Lu. Formulation and efficacy of ECO/pRHO-ABCA4-SV40 nanoparticles for nonviral gene therapy of Stargardt disease in a mouse model. Journal of controlled release : official journal of the Controlled Release Society. 2021 Feb 10;330:329-340