Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
Copernicus Therapeutics, Inc., Cleveland, OH, USA.
Int J Nanomedicine. 2018 Mar 8;13:1361-1379. doi: 10.2147/IJN.S157000. eCollection 2018.
DNA nanoparticles (NPs) comprising polylysine conjugated to polyethylene glycol efficiently target murine photoreceptors and the retinal pigment epithelium (RPE) and lead to long-term phenotypic improvement in models of retinal degeneration. Advancing this technology requires testing in a large animal model, particularly with regard to safety. So, herein we evaluate NPs in non-human primates (baboon).
NPs with plasmids carrying GFP and a ubiquitous, RPE-specific, or photoreceptor-specific promoter were delivered by either subretinal or intravitreal injection. We detected GFP message and protein in the retina/RPE from eyes dosed with NPs carrying ubiquitously expressed and RPE-specific vectors, and GFP message in eyes injected with NPs carrying photoreceptor-specific vectors. Importantly, we observed NP DNA in the retina/RPE following intravitreal injection, indicating the inner limiting membrane does not prevent NP diffusion into the outer retina. We did not observe any adverse events in any baboon, and there were no NP-associated changes in retinal function. Furthermore, no systemic or local inflammatory reaction to the vectors/injections was observed, and no NP DNA was found outside the eye.
Taken together with the well-established rodent safety and efficacy data, these findings suggest that DNA NPs may be a safe and potentially clinically viable nonviral ocular therapy platform for retinal diseases.
由聚赖氨酸与聚乙二醇偶联而成的 DNA 纳米颗粒(NPs)可有效地靶向小鼠感光细胞和视网膜色素上皮(RPE),并在视网膜变性模型中导致长期表型改善。为了推进这项技术,需要在大型动物模型中进行测试,特别是在安全性方面。因此,本文在非人类灵长类动物(狒狒)中评估了 NPs。
通过视网膜下或玻璃体内注射,将携带 GFP 和普遍表达、RPE 特异性或光感受器特异性启动子的质粒的 NPs 递送至眼睛。我们从用携带普遍表达和 RPE 特异性载体的 NPs 处理的眼睛中检测到 GFP 信使和蛋白质在视网膜/RPE 中,从用携带光感受器特异性载体的 NPs 处理的眼睛中检测到 GFP 信使。重要的是,我们在玻璃体内注射后观察到 NP DNA 在视网膜/RPE 中,表明内界膜不能阻止 NP 扩散到外视网膜。在任何一只狒狒中,我们都没有观察到任何不良反应,视网膜功能也没有 NP 相关的变化。此外,没有观察到载体/注射引起的全身或局部炎症反应,也没有发现 NP DNA 出现在眼睛外部。
综上所述,与已建立的啮齿动物安全性和功效数据一起,这些发现表明 DNA NPs 可能是一种安全且具有潜在临床应用价值的非病毒眼部治疗平台,可用于治疗视网膜疾病。