Department of Zoology, University of Oxford, Oxford, OX1 3PS, UK.
Departament de Genètica, Microbiologia i Estadística, Universitat de Barcelona, 08028 Barcelona, Spain.
Genes (Basel). 2019 Apr 9;10(4):289. doi: 10.3390/genes10040289.
Many rare diseases course with affectation of neurosensory organs. Among them, the neuroepithelial retina is very vulnerable due to constant light/oxidative stress, but it is also the most accessible and amenable to gene manipulation. Currently, gene addition therapies targeting retinal tissue (either photoreceptors or the retinal pigment epithelium), as a therapy for inherited retinal dystrophies, use adeno-associated virus (AAV)-based approaches. However, efficiency and safety of therapeutic strategies are relevant issues that are not always resolved in virus-based gene delivery and alternative methodologies should be explored. Based on our experience, we are currently assessing the novel physical properties at the nanoscale of inorganic gold nanoparticles for delivering genes to the retinal pigment epithelium (RPE) as a safe and efficient alternative approach. In this work, we present our preliminary results using DNA-wrapped gold nanoparticles (DNA-gold NPs) for successful in vitro gene delivery on human retinal pigment epithelium cell cultures, as a proof-of-principle to assess its feasibility for retina in vivo gene delivery. Our results show faster expression of a reporter gene in cells transfected with DNA-gold NPs compared to DNA-liposome complexes. Furthermore, we show that the DNA-gold NPs follow different uptake, internalization and intracellular vesicle trafficking routes compared to pristine NPs.
许多罕见病会影响神经感觉器官。其中，神经上皮视网膜由于持续的光/氧化应激而非常脆弱，但它也是最容易接近和适合基因操作的。目前，针对视网膜组织（光感受器或视网膜色素上皮）的基因添加疗法作为遗传性视网膜营养不良的治疗方法，采用基于腺相关病毒（AAV）的方法。然而，治疗策略的效率和安全性是基于病毒的基因传递中尚未解决的相关问题，应该探索替代方法。基于我们的经验，我们目前正在评估无机金纳米粒子在纳米尺度下的新型物理特性，以将基因递送到视网膜色素上皮（RPE）作为一种安全有效的替代方法。在这项工作中，我们使用 DNA 包裹的金纳米粒子（DNA-金 NPs）展示了在人视网膜色素上皮细胞培养物上成功进行体外基因传递的初步结果，作为评估其在体内视网膜基因传递可行性的初步验证。我们的结果表明，与 DNA-脂质体复合物相比，用 DNA-金 NPs 转染的细胞中报告基因的表达更快。此外，我们还表明，与原始 NPs 相比，DNA-金 NPs 具有不同的摄取、内化和细胞内囊泡运输途径。
