Cationic lipid polymerization as a novel approach for constructing new DNA delivery agents.
Cationic lipid polymerization as a novel approach for constructing new DNA delivery agents.
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DOI：
10.1021/bc000097e
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发表时间：
2001-03
影响因子：
4.7
通讯作者：
J. Wu;M. Lizarzaburu;M. Kurth;L. Liu;H. Wege;M. Zern;M. Nantz
中科院分区：
文献类型：
--
作者：
J. Wu;M. Lizarzaburu;M. Kurth;L. Liu;H. Wege;M. Zern;M. Nantz
阳离子脂质介导的体内基因递送通常会因与血液中蛋白质的复合而发生聚集而受到损害。为了提高阳离子脂质-DNA复合物的稳定性，本研究旨在开发一种利用聚阳离子脂质（PCL）形成稳定的阳离子复合物用于基因转染的新方法。β AE-DMRI（PCL的可聚合前体）的丙烯酰胺类似物的氢化提供了单体脂质衍生物（MHL），其用于直接比较相应的脂质复合物稳定性、毒性和转染活性。在本研究中产生并检查了阳离子脂质体的各种制剂，例如MHL、MHL-胆固醇（Chol）、PCL、PCL-Chol、DOTAP-Chol和市售的脂质体。新的聚（阳离子脂质）对大鼠肝细胞或Hep G2细胞没有显示出任何显著的毒性，如LDH泄漏试验所示。此外，PCL的毒性显著低于MHL、DOTAP-Chol或lipofectamine。PCL的悬浮液抗聚集，即使在24小时后暴露于含有50%和100%胎牛血清（FBS）的解决方案。与此相反，脂质体的悬浮液广泛聚集后，24小时的暴露于50%FBS。为了检测脂质聚合对基因转移活性的影响，在Hep G2和亚历山大细胞系中进行荧光素酶载体（pGL 3）的脂质体介导的转染。PCL制剂在Hep G2细胞中的荧光素酶活性与MHL、DOTAP-Chol和脂质体制剂的荧光素酶活性相似，表明脂质聚合不损害转染活性。与单体前体MHL和工业转染标准DOTAP和lipofectamine相比，新型聚（阳离子脂质）表现出最低的细胞毒性，对血清诱导的聚集最有抵抗力，并且当与胆固醇共配制时具有相当的转染活性。这种新的聚合方法的发展稳定和积极的聚合物可能证明是一个有价值的替代体内基因传递。
In vivo gene delivery mediated by cationic lipids is often compromised by aggregation due to complexation with proteins in the blood. To improve the stability of cationic lipid-DNA complexes, the present study aimed to develop a novel approach in which a poly(cationic lipid) (PCL) is utilized to form stable cationic polyplexes for gene transfection. Hydrogenation of the acrylamide analogue of betaAE-DMRI, the polymerizable precursor of PCL, provided a monomeric lipid derivative (MHL) which was used for direct comparison of corresponding lipoplex stability, toxicity, and transfection activity. Various formulations of cationic liposomes, such as MHL, MHL-cholesterol (Chol), PCL, PCL-Chol, DOTAP-Chol, and commercially available lipofectamine were generated and examined in this study. The new poly(cationic lipid) did not display any significant toxicity to rat hepatocytes or Hep G2 cells as indicated by an LDH leakage assay. Furthermore, PCL was significantly less toxic than MHL, DOTAP-Chol or lipofectamine. Suspensions of PCL were resistant to aggregation even after 24 h of exposure to solutions containing 50 and 100% fetal bovine serum (FBS). In contrast, suspensions of lipofectamine extensively aggregated after 24 h of exposure to 50% FBS. To examine the influence of lipid polymerization on gene transfer activity, liposome-mediated transfections of a luciferase vector (pGL3) were performed in Hep G2 and Alexander cell lines. The luciferase activity of the PCL formulations in Hep G2 cells were similar to those of the MHL, DOTAP-Chol and lipofectamine formulations, demonstrating that lipid polymerization does not compromise transfection activity. In comparison to the monomeric precursor MHL and to the industry transfection standards DOTAP and lipofectamine, the novel poly(cationic lipid) exhibited the lowest cytotoxicity, was the most resistant to serum-induced aggregation and had comparable transfection activity when coformulated with cholesterol. This novel polymerization approach for the development of stable and active polyplexes may prove a valuable alternative for in vivo gene delivery.