Targeted Multifunctional Lipid ECO Plasmid DNA Nanoparticles as Efficient Non-viral Gene Therapy for Leber’s Congenital Amaurosis

• Published in: Molecular therapy. Nucleic acids Vol. 7; no. C; pp. 42 - 52
• Main Authors: Sun, Da, Sahu, Bhubanananda, Gao, Songqi, Schur, Rebecca M., Vaidya, Amita M., Maeda, Akiko, Palczewski, Krzysztof, Lu, Zheng-Rong
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16-06-2017; Elsevier Limited; American Society of Gene & Cell Therapy; Elsevier
• Subjects: Blindness; Chemical bonds; Congenital diseases; Deoxyribonucleic acid; DNA; Efficiency; Electrodes; Epithelium; Eye diseases; Gene expression; gene replacement therapy; Gene therapy; Gene transfer; Injection; Interphotoreceptor retinoid-binding protein; Leber’s congenital amaurosis; Lipids; Microscopy; multifunctional lipid; Nanoparticles; Non-viral gene delivery; Original; pH effects; Photoreceptors; Plasmids; Polyethylene glycol; Proteins; Retina; Retinal pigment epithelium; Retinoid-binding protein; RPE65 protein; Transfection; Vectors (Biology); multifunctional lipid; gene replacement therapy; RPE65 protein; Non-viral gene delivery; Leber’s congenital amaurosis
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1016/j.omtn.2017.02.005

Development of a gene delivery system with high efficiency and a good safety profile is essential for successful gene therapy. Here we developed a targeted non-viral delivery system using a multifunctional lipid ECO for treating Leber’s congenital amaurosis type 2 (LCA2) and tested this in a mouse model. ECO formed stable nanoparticles with plasmid DNA (pDNA) at a low amine to phosphate (N/P) ratio and mediated high gene transfection efficiency in ARPE-19 cells because of their intrinsic properties of pH-sensitive amphiphilic endosomal escape and reductive cytosolic release (PERC). All-trans-retinylamine, which binds to interphotoreceptor retinoid-binding protein (IRBP), was incorporated into the nanoparticles via a polyethylene glycol (PEG) spacer for targeted delivery of pDNA into the retinal pigmented epithelium. The targeted ECO/pDNA nanoparticles provided high GFP expression in the RPE of 1-month-old Rpe65−/− mice after subretinal injection. Such mice also exhibited a significant increase in electroretinographic activity, and this therapeutic effect continued for at least 120 days. A safety study in wild-type BALB/c mice indicated no irreversible retinal damage following subretinal injection of these targeted nanoparticles. All-trans-retinylamine-modified ECO/pDNA nanoparticles provide a promising non-viral platform for safe and effective treatment of RPE-specific monogenic eye diseases such as LCA2.
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