Megalin-targeted enhanced transfection efficiency in cultured human HK-2 renal tubular proximal cells using aminoglycoside-carboxyalkyl- polyethylenimine -containing nanoplexes.
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-PEI25(25kDa PEI) or carboxyalkylated-PEI10(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-PEI25and ∼7-fold increase for aminoglycoside-carboxyalkylated-PEI10relative 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.
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.
Fatemeh
F
Oroojalian
Nanobiotechnology Group, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran. Electronic address: ahrezayan@ut.ac.ir.
Ali Hossein
AH
Rezayan
Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, USA.
Wayne Thomas
WT
Shier
Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Khalil
K
Abnous
Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: ramezanim@mums.ac.ir.
Mohammad
M
Ramezani
D000617:Aminoglycosides / Q000008:administration & dosage* / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry; D004285:Dogs; D049452:Green Fluorescent Proteins / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology; D026561:Low Density Lipoprotein Receptor-Related Protein-2 / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D008807:Mice, Inbred BALB C; D049329:Nanostructures / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids; D011094:Polyethyleneimine / Q000008:administration & dosage* / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D000617:Aminoglycosides; D026561:Low Density Lipoprotein Receptor-Related Protein-2; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA
Aminoglycosides; Gene Delivery; Megalin; Polyethylenimine; Renal Targeting