Wang Dan, Wang Xuelei, Wang Li, Zhang Jin, Ma Jie, Xia Guimin, Hong Bin
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China; NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China.
Eur J Pharm Sci. 2021 Jun 1;161:105803. doi: 10.1016/j.ejps.2021.105803. Epub 2021 Mar 13.
MicroRNA185 (miR185), an endogenous noncoding RNA with 23 nucleotides, is one of key posttranscriptional modulators of cholesterol metabolism in hepatic cells. The antisense inhibitor of miR185 (miR185i) could decrease cholesterol level in vivo, providing a promising agent for anti-atherosclerosis strategy. In this work, a novel LipomiR185i was constructed by thin film hydration method and post-PEGylation as DOPE: DOTAP: Chol: DSPE-PEG at the molar ratio of 1:1:1:0.1 with a nitrogen-to-phosphate ratio of 3, through the optimization of three cationic lipids (DOTAP, DODMA and DLin-MC3-DMA), six helper lipids (PC-98T, HSPC, DOPE, DMPC, DPPC and DSPC), different amounts and incorporation approaches of DSPE-PEG and nitrogen-to-phosphate ratio. LipomiR185i was characterized with a particle size of 174 ± 11 nm, a zeta potential of 7.0 ± 3.3 mV, high encapsulation efficiency and transfection activity. It could protect miR185i from the rapid degradation by nucleases in serum, enhance cellular uptake and promote lysosomal escape in HepG2 cells. LipomiR185i could accumulate in the liver and remain for at least two weeks. More importantly, LipomiR185i significantly down-regulated the hepatic endogenous miR185 level in vitro and in vivo without significant tissue damage at 14 mg⋅kg. The construction of LipomiR185i provides a potential anti-atherosclerotic nanodrug as well as a platform for delivering small RNAs to the liver efficiently and safely.
微小RNA185（miR185）是一种含23个核苷酸的内源性非编码RNA，是肝细胞胆固醇代谢关键的转录后调节因子之一。miR185的反义抑制剂（miR185i）可降低体内胆固醇水平，为抗动脉粥样硬化策略提供了一种有前景的药物。在本研究中，通过薄膜水化法和聚乙二醇化后，以1:1:1:0.1的摩尔比将二油酰磷脂酰乙醇胺（DOPE）、二油酰基三甲基氯化铵（DOTAP）、胆固醇（Chol）和二硬脂酰磷脂酰乙醇胺-聚乙二醇（DSPE-PEG）构建了一种新型脂质体miR185i（LipomiR185i），氮磷比为3，通过优化三种阳离子脂质（DOTAP、二油酰基二甲基铵（DODMA）和二油酰基丙基三甲基氯化铵（DLin-MC3-DMA））、六种辅助脂质（PC-98T、氢化大豆卵磷脂（HSPC）、DOPE、二肉豆蔻酰磷脂酰胆碱（DMPC）、二棕榈酰磷脂酰胆碱（DPPC）和二硬脂酰磷脂酰胆碱（DSPC））、DSPE-PEG的不同用量和掺入方法以及氮磷比。LipomiR185i的粒径为174±11nm，zeta电位为7.0±3.3mV，具有高包封率和转染活性。它可以保护miR185i不被血清中的核酸酶快速降解，增强细胞摄取并促进其在HepG2细胞中的溶酶体逃逸。LipomiR185i可在肝脏中蓄积并至少保留两周。更重要的是，LipomiR185i在体外和体内均能显著下调肝脏内源性miR185水平，在剂量为14mg·kg时无明显组织损伤。LipomiR185i的构建提供了一种潜在的抗动脉粥样硬化纳米药物以及一个将小RNA高效安全地递送至肝脏的平台。
