Direct gene transfer with compacted DNA nanoparticles in retinal pigment epithelial cells: expression, repeat delivery and lack of toxicity. (April 2012)

• Record Type: Journal Article
• Authors: Han, Zongchao; Koirala, Adarsha; Makkia, Rasha; Cooper, Mark J; Naash, Muna I
• Is Part Of: Nanomedicine. Volume 7:Number 4(2012)
• Page Start: 521
• Page End: 539
• Publication Date: 2012-04
• Subjects: enhanced green fluorescent protein -- gene transfer -- nanoparticle -- nonviral -- repeat delivery -- retinal pigment epithelial cell -- subretinal injection -- toxicity; Nanotechnology -- Periodicals; Medical technology -- Periodicals; Nanotechnology -- Therapeutic use -- Periodicals; 610.28
• DOI: 10.2217/nnm.11.158
• Languages: English
• ISSNs: 1743-5889

Abstract

Aim: To evaluate the safety of compacted DNA nanoparticles (NPs) in retinal pigment epithelial (RPE) cells.Materials & methods: Enhanced GFP expression cassettes controlled by the RPE-specific vitelloform macular dystrophy promoter were constructed with and without a bacterial backbone and compacted into NPs formulated with polyethylene glycol-substituted lysine 30-mers. Single or double subretinal injections were administered in adult BALB/c mice. Expression levels of enhanced GFP, proinflammatory cytokines and neutrophil/macrophage mediators, and retinal function by electroretinogram were evaluated at different time-points postinjection.Results: Immunohistochemistry and real-time PCR demonstrated that NPs specifically transfect RPE cells at a higher efficiency than naked DNA and similar results were observed after the second injection. At 6 h postinjections, a transient inflammatory response was observed in all cohorts, including saline, indicating an adverse effect to the injection procedure. Subsequently, no inflammation was detected in all experimental groups.Conclusion: This study demonstrates the safety and efficacy of NP-mediated RPE gene transfer therapy following multiple subretinal administrations.