Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy.
Department of Drug Science and Technology, University of Turin, Via Pietro Giuria, 11, 10125, Turin, Italy.
Nanomedicine. 2022 Sep;45:102589. doi: 10.1016/j.nano.2022.102589. Epub 2022 Jul 28.
Design of nanocarriers for efficient miRNA delivery can significantly improve miRNA-based therapies. Lipoplexes based on helper lipid, dioleoyl phosphatidylethanolamine (DOPE) and cationic lipid [2-(2,3-didodecyloxypropyl)-hydroxyethyl] ammonium bromide (DE) were formulated to efficiently deliver miR-1 or a combination of four microRNAs (miRcombo) to adult human cardiac fibroblasts (AHCFs). Lipoplexes with amino-to-phosphate groups ratio of 3 (N/P 3) showed nanometric hydrodynamic size (372 nm), positive Z-potential (40 mV) and high stability under storage conditions. Compared to commercial DharmaFECT1 (DF), DE-DOPE/miRNA lipoplexes showed superior miRNA loading efficiency (99 % vs. 64 %), and faster miRNA release (99 % vs. 82 % at 48 h). DE-DOPE/miR-1 lipoplexes showed superior viability (80-100 % vs. 50 %) in AHCFs, a 2-fold higher miR-1 expression and Twinfilin-1 (TWF-1) mRNA downregulation. DE-DOPE/miRcombo lipoplexes significantly enhanced AHCFs reprogramming into induced cardiomyocytes (iCMs), as shown by increased expression of CM markers compared to DF/miRcombo.
纳米载体的设计可有效提高 miRNA 的传递效率，从而显著改善 miRNA 疗法。本文构建了基于辅助脂质二油酰基磷脂酰乙醇胺（DOPE）和阳离子脂质[2-(2,3-二二十二烷氧基丙基)-羟乙基]溴化铵（DE）的阳离子脂质体，用于有效递送至成年人心房成纤维细胞（AHCFs）的 miR-1 或四种 microRNA 混合物（miRcombo）。具有氨基与磷酸基团比例为 3（N/P 3）的阳离子脂质体具有纳米级流体力学粒径（372nm）、正 zeta 电位（40mV），且在储存条件下具有高稳定性。与商业试剂 DharmaFECT1（DF）相比，DE-DOPE/miRNA 脂质体表现出更高的 miRNA 负载效率（99%对 64%）和更快的 miRNA 释放（48h 时 99%对 82%）。DE-DOPE/miR-1 脂质体在 AHCFs 中表现出更好的细胞活力（80-100%对 50%），miR-1 表达更高，Twinfilin-1（TWF-1）mRNA 下调 2 倍。与 DF/miRcombo 相比，DE-DOPE/miRcombo 脂质体显著增强了 AHCFs 向诱导型心肌细胞（iCM）的重编程，表现为 CM 标志物表达增加。
