Oroojalian Fatemeh, Rezayan Ali Hossein, Shier Wayne Thomas, Abnous Khalil, Ramezani Mohammad
Nanobiotechnology Group, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran; Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, USA.
Nanobiotechnology Group, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Int J Pharm. 2017 May 15;523(1):102-120. doi: 10.1016/j.ijpharm.2017.03.024. Epub 2017 Mar 16.
Non-viral vectors are of interest as therapeutic gene delivery agents in gene therapy, because they are simple to prepare, easy to modify and have definable safety profiles compared to viral vectors. The potential of gene therapy in the treatment of renal diseases is limited by a lack of effective kidney-targeted gene delivery systems. Aminoglycoside antibiotics gentamicin and neomycin were connected by amide linkages to carboxyl groups on carboxyalkylated-PEI (25kDa PEI) or carboxyalkylated-PEI (10kDa PEI). Aminoglycoside-carboxyalkylated-PEI conjugates were characterized with respect to size, surface charge density, DNA condensation ability, and buffering capacity. Polyplexes prepared by electrostatic interaction between aminoglycoside-carboxyalkylated-PEIs and enhanced green fluorescent protein-expressing (EGFP) plasmid DNA had appropriate nano-scale size (143-173nm). Their targeting potential was investigated in cultured HK-2 immortalized human cortex/proximal tubule kidney epithelial cells, which expresses megalin, a scavenger receptor that mediates endocytosis of a diverse group of ligands, including aminoglycoside antibiotics. Aminoglycoside-carboxyalkylated-PEIs significantly increased EGFP gene transfection efficiency in HK-2 cells by ∼13-fold for aminoglycoside-carboxyalkylated-PEI and ∼7-fold increase for aminoglycoside-carboxyalkylated-PEI relative to the corresponding PEIs without aminoglycosides. The transfection efficiency of polyplexes was dependent on the weight ratio of aminoglycoside-containing ligand in the carrier. In the presence of a range of concentrations of human serum albumin, which competes for megalin binding, aminoglycoside-carboxyalkylated-PEI-mediated transfection was reduced to background levels. These results suggest that aminoglycoside-carboxyalkylated-PEI polyplexes can target megalin-expressing kidney-derived cells in vitro resulting in improved transfection efficiency with low cytotoxicity.
在基因治疗中，非病毒载体作为治疗性基因递送剂备受关注，因为与病毒载体相比，它们易于制备、易于修饰且具有明确的安全性。基因治疗在肾脏疾病治疗中的潜力因缺乏有效的肾脏靶向基因递送系统而受到限制。氨基糖苷类抗生素庆大霉素和新霉素通过酰胺键与羧烷基化聚乙二醇（25kDa聚乙二醇）或羧烷基化聚乙二醇（10kDa聚乙二醇）上的羧基相连。对氨基糖苷-羧烷基化聚乙二醇缀合物的大小、表面电荷密度、DNA凝聚能力和缓冲能力进行了表征。通过氨基糖苷-羧烷基化聚乙二醇与增强型绿色荧光蛋白表达（EGFP）质粒DNA之间的静电相互作用制备的多聚体具有合适的纳米级尺寸（143-173nm）。在培养的HK-2永生化人皮质/近端肾小管肾上皮细胞中研究了它们的靶向潜力，该细胞表达巨蛋白，一种介导多种配体内吞作用的清道夫受体，包括氨基糖苷类抗生素。与不含氨基糖苷的相应聚乙二醇相比，氨基糖苷-羧烷基化聚乙二醇使HK-2细胞中的EGFP基因转染效率显著提高，氨基糖苷-羧烷基化聚乙二醇提高了约13倍，氨基糖苷-羧烷基化聚乙二醇提高了约7倍。多聚体的转染效率取决于载体中含氨基糖苷配体的重量比。在一系列浓度的人血清白蛋白存在下，人血清白蛋白竞争巨蛋白结合，氨基糖苷-羧烷基化聚乙二醇介导的转染降低至背景水平。这些结果表明，氨基糖苷-羧烷基化聚乙二醇多聚体可以在体外靶向表达巨蛋白的肾源性细胞，从而提高转染效率且细胞毒性低。
