Ko Tomotaka, Fumoto Shintaro, Kurosaki Tomoaki, Nakashima Moe, Miyamoto Hirotaka, Sasaki Hitoshi, Nishida Koyo
Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Pharmaceutics. 2024 Apr 9;16(4):522. doi: 10.3390/pharmaceutics16040522.
Typical examples of non-viral vectors are binary complexes of plasmid DNA with cationic polymers such as polyethyleneimine (PEI). However, problems such as cytotoxicity and hemagglutination, owing to their positively charged surfaces, hinder their in vivo use. Coating binary complexes with anionic polymers, such as γ-polyglutamic acid (γ-PGA), can prevent cytotoxicity and hemagglutination. However, the role of interactions between these complexes and serum components in in vivo gene transfer remains unclear. In this study, we analyzed the contribution of serum components to in vivo gene transfer using PEI/plasmid DNA binary complexes and γ-PGA/PEI/plasmid DNA ternary complexes. In binary complexes, heat-labile components in the serum greatly contribute to the hepatic and splenic gene expression of the luciferase gene. In contrast, serum albumin and salts affected the hepatic and splenic gene expression in the ternary complexes. Changes in physicochemical characteristics, such as increased particle size and decreased absolute values of ζ-potential, might be involved in the enhanced gene expression. These findings would contribute to a better understanding of in vivo non-viral gene transfer using polymers, such as PEI and γ-PGA.
非病毒载体的典型例子是质粒DNA与阳离子聚合物（如聚乙烯亚胺，PEI）形成的二元复合物。然而，由于其带正电的表面，诸如细胞毒性和血凝等问题阻碍了它们在体内的应用。用阴离子聚合物（如γ-聚谷氨酸，γ-PGA）包覆二元复合物可以防止细胞毒性和血凝。然而，这些复合物与血清成分之间的相互作用在体内基因转移中的作用仍不清楚。在本研究中，我们使用PEI/质粒DNA二元复合物和γ-PGA/PEI/质粒DNA三元复合物分析了血清成分对体内基因转移的贡献。在二元复合物中，血清中的热不稳定成分对荧光素酶基因的肝脏和脾脏基因表达有很大贡献。相反，血清白蛋白和盐影响三元复合物中的肝脏和脾脏基因表达。诸如粒径增加和ζ-电位绝对值降低等物理化学特性的变化可能与基因表达增强有关。这些发现将有助于更好地理解使用PEI和γ-PGA等聚合物进行的体内非病毒基因转移。
