Department of Pharmaceutical Sciences, School of Pharmacy, College of Health Professions, North Dakota State University, Fargo 58105, ND, USA.
Department of Pharmaceutical Sciences, School of Pharmacy, College of Health Professions, North Dakota State University, Fargo 58105, ND, USA.
Int J Pharm. 2021 Oct 25;608:121095. doi: 10.1016/j.ijpharm.2021.121095. Epub 2021 Sep 20.
Vgf (non-acronymic), a neurotrophin stimulated protein which plays a crucial role in learning, synaptic activity, and neurogenesis, is markedly downregulated in the brain of Alzheimer's disease (AD) patients. However, since vgf is a large polar protein, a safe and efficient gene delivery vector is critical for its delivery across the blood brain barrier (BBB). This research work demonstrates brain-targeted liposomal nanoparticles optimized for delivering plasmid encoding vgf across BBB and transfecting brain cells. Brain targeting was achieved by surface functionalization using glucose transporter-1 targeting ligand (mannose) and brain targeted cell-penetrating peptides (chimeric rabies virus glycoprotein fragment, rabies virus derived peptide, penetratin peptide, or CGNHPHLAKYNGT peptide). The ligands were conjugated to lipid via nucleophilic substitution reaction resulting in >75% binding efficiency. The liposomes were formed by film hydration technique demonstrating size <200 nm, positive zeta potential (15-20 mV), and polydispersity index <0.3. The bifunctionalized liposomes demonstrated ∼3 pg/µg protein vgf transfection across in vitro BBB, and ∼80 pg/mg protein in mice brain which was 1.5-2 fold (p < 0.05) higher compared to untreated control. The nanoparticles were also biocompatible in vitro and in vivo, suggesting a safe and efficient gene delivery system to treat AD.
Vgf（非首字母缩写词）是一种神经营养因子刺激蛋白，在学习、突触活动和神经发生中起着至关重要的作用，在阿尔茨海默病（AD）患者的大脑中显著下调。然而，由于 vgf 是一种大型极性蛋白，因此安全有效的基因传递载体对于其穿过血脑屏障（BBB）至关重要。这项研究工作展示了经过优化的脑靶向脂质体纳米颗粒，可用于将编码 vgf 的质粒递送到 BBB 并转染脑细胞。通过使用葡萄糖转运蛋白-1 靶向配体（甘露糖）和脑靶向细胞穿透肽（嵌合狂犬病病毒糖蛋白片段、狂犬病病毒衍生肽、穿透肽或 CGNHPHLAKYNGT 肽）进行表面功能化来实现脑靶向。配体通过亲核取代反应与脂质结合，结合效率>75%。通过薄膜水化技术形成脂质体，其粒径<200nm，正zeta 电位（15-20mV），多分散指数<0.3。双功能化脂质体在体外 BBB 中显示出3pg/μg 蛋白 vgf 转染，在小鼠脑中为80pg/mg 蛋白，与未处理的对照组相比提高了 1.5-2 倍（p<0.05）。该纳米颗粒在体外和体内也具有生物相容性，表明这是一种安全有效的基因传递系统，可用于治疗 AD。
