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

基因治疗的成功在很大程度上取决于递送载体或载体的开发，其可以选择性地和有效地将治疗基因递送到靶细胞，并且毒性最小。本研究的目的是发展基于系统控制递送和转染的体内基因递送优化策略。首先，开发了用于pDNA的新型残基化放射性标记。将4-[对叠氮基水杨酰胺基]丁胺（ASBA）与二乙烯三胺五乙酸（DTPA）酸酐偶联，经光活化后与pDNA反应，再用[^<111>In]InCl_3标记得到^<111>In-pDNA。这表明，μ <111>In-pDNA不仅可用于评估pDNA及其与载体系统的复合物的组织分布，而且可用于设计用于靶向递送pDNA的有效载体。就体内选择性基因传递至肝细胞而言，半乳糖已被证明是一种有前途的靶向配体 ...更多信息 肝细胞（肝实质细胞），因为该细胞具有大量的去唾液酸糖蛋白受体，其识别糖蛋白的寡糖链上或化学合成的半乳糖基化载体上的半乳糖单位。我们假设，在脂质复合物形成过程中存在必需量的氯化钠（NaCl）可能会调节阳离子脂质体之间的排斥，阳离子脂质体在脂质复合物中的融合将通过部分中和正电荷而加速。FRET分析表明，脂质复合物中的NaCl溶液减弱了阳离子脂质体之间的排斥作用，增强了阳离子脂质体在脂质复合物中的融合，从而大大增强了在肝细胞中的体内转染。S-亚硝基硫醇是一类有趣的一氧化氮（NO）供体，用于治疗循环障碍。在这项研究中，我们开发了一种新的大分子NO供体，其中10个NO分子通过S-亚硝基硫醇键共价键合到聚乙二醇（PEG）缀合的牛血清白蛋白（BSA）（PEG-poly SNO-BSA）。这表明，新的S-亚硝基硫醇PEG-聚SNO-BSA是一个有前途的化合物，表现出独特的特性，持续释放NO在体内血液循环中，这将是有益的治疗循环障碍。这些信息将是有价值的非病毒载体的临床应用的发展。少

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