Kurosaki Tomoaki, Kanda Hiroki, Hashizume Junya, Sato Kayoko, Harasawa Hitomi, Nakamura Tadahiro, Sasaki Hitoshi, Kodama Yukinobu
Department of Hospital Pharmacy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8588, Japan.
Pharmaceutics. 2021 Nov 22;13(11):1983. doi: 10.3390/pharmaceutics13111983.
In a previous study, we constructed a lung-targeting lipopolyplex containing polyethyleneimine (PEI), 1,2-di--octadecenyl-3-trimethylammonium propane (DOTMA), and -lauroylsarcosine (LS). The lipopolyplex exhibited an extremely high gene expression in the lung after intravenous administration. Here, we optimized the lipopolyplex and used it to deliver a TGF-β1 shRNA to treat refractory pulmonary fibrosis. We constructed several lipopolyplexes with pDNA, various cationic polymers, cationic lipids, and LS to select the most effective formulation. Then, the pDNA encoding shRNA against mouse TGF-β1 was encapsulated in the lipopolyplex and injected into mice with bleomycin-induced pulmonary fibrosis. After optimizing the lipopolyplex, dendrigraft poly-L-lysine (DGL) and DOTMA were selected as the appropriate cationic polymer and lipid, respectively. The lipopolyplex was constructed with a pDNA, DGL, DOTMA, and LS charge ratio of 1:2:2:4 showed the highest gene expression. After intravenous administration of the lipopolyplex, the highest gene expression was observed in the lung. In the in vitro experiment, the lipopolyplex delivered pDNA into the cells via endocytosis. As a result, the lipopolyplex containing pDNA encoding TGF-β1 shRNA significantly decreased hydroxyproline in the pulmonary fibrosis model mice. We have successfully inhibited pulmonary fibrosis using a novel lung-targeting lipopolyplex.
在先前的一项研究中，我们构建了一种肺靶向脂质多聚体，其包含聚乙烯亚胺（PEI）、1,2-二油酰基-3-三甲基铵丙烷（DOTMA）和月桂酰肌氨酸（LS）。静脉注射后，该脂质多聚体在肺中表现出极高的基因表达。在此，我们对脂质多聚体进行了优化，并使用其递送转化生长因子-β1（TGF-β1）短发夹RNA（shRNA）来治疗难治性肺纤维化。我们用质粒DNA（pDNA）、各种阳离子聚合物、阳离子脂质和LS构建了几种脂质多聚体，以选择最有效的配方。然后，将编码针对小鼠TGF-β1的shRNA的pDNA封装在脂质多聚体中，并注射到博来霉素诱导的肺纤维化小鼠体内。在优化脂质多聚体后，分别选择树枝状聚-L-赖氨酸（DGL）和DOTMA作为合适的阳离子聚合物和脂质。以1:2:2:4的pDNA、DGL、DOTMA和LS电荷比构建的脂质多聚体显示出最高的基因表达。静脉注射脂质多聚体后，在肺中观察到最高的基因表达。在体外实验中，脂质多聚体通过内吞作用将pDNA递送至细胞内。结果，含有编码TGF-β1 shRNA的pDNA的脂质多聚体显著降低了肺纤维化模型小鼠中的羟脯氨酸水平。我们已使用新型肺靶向脂质多聚体成功抑制了肺纤维化。
