Strategy for optimization of in vivo gene delivery based on systemic control of delivery and transfection

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The success of gene therapy largely depends upon the development of delivery vehicles or vectors, which can selectively and efficiently deliver therapeutic genes to target cells with minimal toxicity. The purpose of this study was to develop the strategy for optimization of in vivo gene delivery based on systemic control of delivery and transfection. First, a novel residualizing radiolabel for pDNA was developed. 4-[p-Azidosalicylamido]butylamine (ASBA) was coupled with diethylenetriaminepentaacetic acid (DTPA) anhydride, then the conjugate was reacted with pDNA by photoactivation, followed by labeling with [^<111>In]InCl_3 to obtain ^<111>In-pDNA. It is suggested that ^<111>In-pDNA is useful not only for evaluating the tissue distribution of pDNA and its complex with a vector system, but also for designing an effective vector for targeted delivery of pDNA. As far as in vivo selective gene delivery to hepatocytes is concerned, galactose has been shown to be a promising targeting ligand … More to hepatocytes (liver parenchymal cells) because the cells possess a large number of asialoglycoprotein receptors that recognize the galactose units on the oligosaccharide chains of glycoproteins or on the chemically synthesized galactosylated carriers. We hypothesized that the presence of an essential amount of sodium chloride (NaCl) during lipoplex formation might regulate repulsion between cationic liposomes and fusion of cationic liposomes in the lipoplex would be accelerated by partial neutralization of the positive charge. FRET analysis revealed that the NaCl solution in the lipoplex weakened the repulsion among cationic liposomes and enhanced the fusion of cationic liposomes in the lipoplex ; consequently, the in vivo transfection in hepatocytes was greatly enhanced. S-Nitrosothiols are an interesting class of nitric oxide (NO) donors used for the treatment of circulation disorders. In this study, we developed a novel macromolecular NO donor in which 10 NO molecules were covalently bound to polyethylene glycol (PEG)-conjugated bovine serum albumin (BSA) through S-nitrosothiol linkages (PEG-poly SNO-BSA). It is suggested that the novel S-nitrosothiol PEG-poly SNO-BSA is a promising compound that exhibits unique characteristics of sustained release of NO in the blood circulation in vivo, which would be beneficial for the treatment of circulation disorders. This information will be valuable for the development of nonviral vectors for clinical applications. Less