Sen Merve, Bassetto Marco, Poulhes Florent, Zelphati Olivier, Ueffing Marius, Arango-Gonzalez Blanca
Centre of Ophthalmology, Institute for Ophthalmic Research, University of Tübingen, 72076 Tübingen, Germany.
Graduate Training Centre of Neuroscience, University of Tübingen, 72074 Tübingen, Germany.
Pharmaceutics. 2021 Feb 6;13(2):225. doi: 10.3390/pharmaceutics13020225.
The use of synthetic RNA for research purposes as well as RNA-based therapy and vaccination has gained increasing importance. Given the anatomical seclusion of the eye, small interfering RNA (siRNA)-induced gene silencing bears great potential for targeted reduction of pathological gene expression that may allow rational treatment of chronic eye diseases in the future. However, there is yet an unmet need for techniques providing safe and efficient siRNA delivery to the retina. We used magnetic nanoparticles (MNPs) and magnetic force (Reverse Magnetofection) to deliver siRNA/MNP complexes into retinal explant tissue, targeting valosin-containing protein (VCP) previously established as a potential therapeutic target for autosomal dominant retinitis pigmentosa (adRP). Safe and efficient delivery of VCP siRNA was achieved into all retinal cell layers of retinal explants from the RHO P23H rat, a rodent model for adRP. No toxicity or microglial activation was observed. silencing led to a significant decrease of retinal degeneration. Reverse Magnetofection thus offers an effective method to deliver siRNA into retinal tissue. Used in combination with retinal organotypic explants, it can provide an efficient and reliable preclinical test platform of RNA-based therapy approaches for ocular diseases.
合成RNA在研究目的以及基于RNA的治疗和疫苗接种中的应用变得越来越重要。鉴于眼睛的解剖学隔离，小干扰RNA（siRNA）诱导的基因沉默对于靶向降低病理性基因表达具有巨大潜力，这可能在未来实现对慢性眼病的合理治疗。然而，目前仍需要一种能够将siRNA安全有效地递送至视网膜的技术。我们使用磁性纳米颗粒（MNP）和磁力（反向磁转染）将siRNA/MNP复合物递送至视网膜外植体组织，靶向含缬酪肽蛋白（VCP），该蛋白先前已被确立为常染色体显性遗传性视网膜色素变性（adRP）的潜在治疗靶点。在RHO P23H大鼠（一种adRP的啮齿动物模型）的视网膜外植体的所有视网膜细胞层中均实现了VCP siRNA的安全有效递送。未观察到毒性或小胶质细胞激活。基因沉默导致视网膜变性显著减少。因此，反向磁转染提供了一种将siRNA递送至视网膜组织的有效方法。与视网膜器官型外植体联合使用时，它可为眼部疾病的基于RNA的治疗方法提供一个高效且可靠的临床前测试平台。
