Enhancement of Gene Expression by Polyamidoamine Dendrimer Conjugates with α-, β-, and γ-Cyclodextrins

Abstract

To improve the transfection efficiency of nonviral vector, we synthesized the starburst polyamidoamine dendrimer conjugates with alpha-, beta-, and gamma-cyclodextrins (CDE conjugates), expecting the synergistic effect of dendrimer and cyclodextrins (CyDs). The (1)H NMR spectroscopic data indicated that alpha-, beta-, and gamma-CyDs are covalently bound to dendrimer in a molar ratio of 1:1. The agarose gel electrophoretic studies revealed that CDE conjugates formed the complexes with plasmid DNA (pDNA) and protected the degradation of pDNA by DNase I in the same manner as dendrimer. CDE conjugates showed a potent luciferase gene expression, especially in the dendrimer conjugate with alpha-CyD (alpha-CDE conjugate) which provided the greatest transfection activity (approximately 100 times higher than those of dendrimer alone and of the physical mixture of dendrimer and alpha-CyD) in NIH3T3 and RAW264.7 cells. In addition, the gene transfer activity of alpha-CDE conjugate was superior to that of Lipofectin. The enhancing gene transfer effect of alpha-CDE conjugate may be attributable to not only increasing the cellular association, but also changing the intracellular trafficking of pDNA. These findings suggest that alpha-CDE conjugate could be a new preferable nonviral vector of pDNA.

Supramolecular Complex of Methyl-β-cyclodextrin with Adamantane-Grafted Hyaluronic Acid as a Novel Antitumor Agent

“…Herein, we previously reported that CyDs accelerate endosomal escape through the interaction of CyDs with phospholipids and cholesterol. [37][38][39][40][41] Therefore, Ad-HA/M-β-CyD may be also able to escape from endosomes through the interaction between M-β-CyD released from Ad-HA/M-β-CyD and phospholipids or cholesterol of endosomal membranes. Additionally, Ad-HA/M-β-CyD colocalized with rhodamine 123, a mitochondria marker (Fig.…”

Polyethylenimine600-β-cyclodextrin: a promising nanopolymer for nonviral gene delivery of primary mesenchymal stem cells

“…Combined with either viral or nonviral gene carriers, CyDs can enhance the gene-transfer efficiencies of these carriers. [13][14][15] We synthesized PEI 600 -β-CyD, a cationic polymer, in which PEI polymers with an MW of 600 Da are linked by β-CyD with low toxicity and high efficiency in neurons. 16 Bone morphogenetic proteins (BMPs) are vital growth factors in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) during osteogenesis and fracture repair.…”

Enhancing Effects of Galactosylated Dendrimer/.ALPHA.-Cyclodextrin Conjugates on Gene Transfer Efficiency

“…Arima et al previously reported the potential use of polyamidoamine (PAMAM) starburst dendrimer (generation 3, G3) bearing a-CyD with the degree of substitution (DS) of 2.4 because of the highest transfection efficiency in vitro and in vivo with low cytotoxicity. [14][15][16] In addition, Arima et al described that mannosylated a-CDE conjugate with the DS values of 3 of the mannose moiety can enhance gene transfer activity of pDNA in various cells with less cytotoxicity.…”

“…14) Galactose residues were attached to primary amino residues of a-CDE conjugate according to the method of Liang et al 18) Five hundreds ml of sodium chloride solution (0.9%, pH 9.0) containing a-CDE conjugate (45 mg) and dimethyl sulfoxide (DMSO) solution containing 0.5 ml of a-D-galactopyranosylphenyl isothiocyanate (DSG 1; 4 mg; DSG 4; 16 mg; DSG 5; 20 mg; DSG 8, 32 mg; and DSG 15, 60 mg) were mixed at 25°C for 24 h. Gal-a-CDE conjugates were purified by gel-filtration (TOSOH TSKGel HW-40S, Tokyo, Japan). 1 H-NMR spectra of these conjugates were measured, and the molar ratios of a-CyD and galactose residues were calculated from the peak areas of anomeric proton of a-CyD and phenyl protons of the a-D-galactopyranosylphenyl isothiocyanate.…”