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输送载体或载体的发展，它们可以选择性地、高效地将治疗性基因输送到毒性最小的靶细胞。本研究的目的是开发基于系统控制的基因传递和转染的体内基因传递的优化策略。首先，开发了一种新的残存放射性标记的PDNA。4-[对-叠氮水杨酰胺]丁胺(ASBA)与二亚乙三胺五乙酸酐(DTPA)偶联，然后通过光活化与PDNA反应，然后用InCl3标记得到^&lt；111&gt；In-PDNA。提示pDNA内载体不仅可用于评价pDNA及其与载体系统的复合体的组织分布，还可用于设计有效的靶向pDNA载体。就体内选择性基因递送到肝细胞而言，半乳糖已被证明是一种很有前途的靶向配体…更多的是肝细胞(肝实质细胞)，因为细胞拥有大量的去唾液酸糖蛋白受体，识别糖蛋白寡糖链上的半乳糖单元或化学合成的半乳糖化载体上的半乳糖单元。我们推测，在脂复合体形成过程中存在必要数量的氯化钠可能会调节阳离子脂质体之间的排斥力，阳离子脂质体在脂复合体中的融合将通过正电荷的部分中和而加速。FRET分析表明，脂复合体中的氯化钠溶液减弱了阳离子脂质体之间的排斥力，促进了阳离子脂质体在脂复合体中的融合，从而大大提高了肝细胞的体内转染率。S-亚硝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，我们开发了一种新型的大分子NO供体，其中10个NO分子通过S-亚硝硫键共价结合到聚乙二醇偶联牛血清白蛋白上(聚乙二醇聚SNO-牛血清白蛋白)。提示新型的S-亚硝硫醇聚乙二醇聚亚硝基牛血清白蛋白是一种具有独特的体内血液循环中NO缓释特性的化合物，有望用于治疗血液循环障碍。这些信息将对临床应用的非病毒载体的开发有价值。较少

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