Petkova Asya I, Kubajewska Ilona, Vaideanu Alexandra, Schätzlein Andreas G, Uchegbu Ijeoma F
UCL School of Pharmacy, 29-39 Brunswick Square, London WC1N 1AX, UK.
Nanomerics Ltd., Northwick Park and St. Mark's Hospital, Y Block, Watford Road, London HA1 3UJ, UK.
Pharmaceutics. 2022 May 26;14(6):1136. doi: 10.3390/pharmaceutics14061136.
Gene delivery to the cerebral cortex is challenging due to the blood brain barrier and the labile and macromolecular nature of DNA. Here we report gene delivery to the cortex using a glycol chitosan-DNA polyplex (GCP). In vitro, GCPs carrying a reporter plasmid DNA showed approximately 60% of the transfection efficiency shown by Lipofectamine lipoplexes (LX) in the U87 glioma cell line. Aiming to maximise penetration through the brain extracellular space, GCPs were coated with hyaluronidase (HYD) to form hyaluronidase-coated polyplexes (GCPH). The GCPH formulation retained approximately 50% of the in vitro hyaluronic acid (HA) digestion potential but lost its transfection potential in two-dimensional U87 cell lines. However, intranasally administered GCPH (0.067 mg kg DNA) showed high levels of gene expression (IVIS imaging of protein expression) in the brain regions. In a separate experiment, involving GCP, LX and naked DNA, the intranasal administration of the GCP formulation (0.2 mg kg DNA) resulted in protein expression predominantly in the cerebral cortex, while a similar dose of intranasal naked DNA led to protein expression in the cerebellum. Intranasal LX formulations did not show any evidence of protein expression. GCPs may provide a means to target protein expression to the cerebral cortex via the intranasal route.
由于血脑屏障以及DNA的不稳定和大分子性质，将基因递送至大脑皮层具有挑战性。在此，我们报告了使用乙二醇壳聚糖-DNA复合纳米粒（GCP）将基因递送至皮层的情况。在体外，携带报告质粒DNA的GCP在U87胶质瘤细胞系中的转染效率约为Lipofectamine脂质体复合物（LX）的60%。为了最大程度地穿透脑细胞外空间，GCP用透明质酸酶（HYD）包被以形成透明质酸酶包被的复合纳米粒（GCPH）。GCPH制剂保留了约50%的体外透明质酸（HA）消化潜力，但在二维U87细胞系中失去了转染潜力。然而，经鼻给药的GCPH（0.067 mg/kg DNA）在脑区显示出高水平的基因表达（蛋白质表达的IVIS成像）。在另一项涉及GCP、LX和裸DNA的实验中，经鼻给药的GCP制剂（0.2 mg/kg DNA）主要在大脑皮层导致蛋白质表达，而相似剂量的经鼻裸DNA则导致蛋白质在小脑表达。经鼻LX制剂未显示任何蛋白质表达的迹象。GCP可能提供一种通过鼻内途径将蛋白质表达靶向大脑皮层的方法。
