Effect of serum components on the physico-chemical properties of cationic lipid/oligonucleotide complexes and on their interactions with cells
Effect of serum components on the physico-chemical properties of cationic lipid/oligonucleotide complexes and on their interactions with cells
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DOI：
10.1016/s0005-2760(97)00169-0
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发表时间：
1998-02-16
期刊：
影响因子：
--
通讯作者：
Szoka, FC
中科院分区：
文献类型：
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作者：
Zelphati, O;Uyechi, LS;Szoka, FC
血清成分和阳离子脂-核酸复合体之间的相互作用对于理解血清如何在体外和体内抑制寡核苷酸的细胞递送是至关重要的。在这项研究中，我们发现几种血清蛋白，特别是牛血清白蛋白(BSA)、脂蛋白(HDL和LDL)和巨球蛋白，与阳离子脂/寡核苷酸复合体相互作用，改变复合体的直径和Zeta电位(从正值到负值)，并显著干扰1，2-二油酰基-3-三甲基氨丙烷(DOTAP)将硫代寡核苷酸(ODN)运送到细胞内的能力。血清和BSA不分离ODN和脂类成分，因此传递的抑制不能归因于ODN中的阳离子脂类的置换。相反，2.5 mg/ml的BSA，与10%血清中的含量相当，使ODN的细胞结合减少约5倍，核摄取减少20倍以上，相反，另一主要血清成分免疫球蛋白G将Zeta电位从正值改变为近中性，对复合体的直径影响不大，但不影响10%血清中发现的量的ODN的细胞结合或核递送。血清中发现的其他分子，特别是油酸和肝素，会取代复合体中的ODN，从而干扰转运。这种位移通过首先与BSA孵育复合体来减弱。血清-复合体相互作用的另一个表现是ODN显著地与阳离子脂质体轻微地激活补体。然而，复合体的形成显著降低了ODN的补体活性。最后，血清的影响可以通过选择辅助脂质(兴奋剂或胆固醇)来部分抵消。辅助性脂质的加入降低了将ODN输送到细胞中所需的有效电荷比(阳离子基团/阴离子硫酸盐)，并允许在培养液中存在较高百分比的血清的情况下输送。这些结果支持了目前的观点，即血清蛋白与复合体的结合是调节阳离子脂质-ODN复合体在培养和静脉给药后活性的重要因素。(C)1998年爱思唯尔科学公司。
The interactions among serum components and cationic lipid-nucleic acid complexes are central to the understanding of how serum inhibits cellular delivery of oligonucleotides in vitro and in vivo. In this study, we show that several serum proteins, in particular bovine serum albumin (BSA), lipoproteins (HDL and LDL) and macroglobulin, interact with cationic lipid/oligonucleotide complexes, alter the complex diameter and zeta potential (from positive to negative values), and significantly interfere with the ability of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) to deliver phosphorothioate oligonucleotides (ODN) into cells. Serum and BSA do not dissociate the ODN and lipid components, therefore inhibition of delivery cannot hp attributed tn a displacement nf cationic lipid from the ODN. Rather BSA at 2.5mg/ml, comparable to the amount found in 10% serum, decreases the cell association of ODN by about 5-fold and nuclear uptake of ODN by greater than 20-fold, In contrast, immunoglobulin G, the other major serum component, alters the zeta potential from positive to near neutral, has a modest effect on the diameter of the complex but does not affect cell association or nuclear delivery of the ODN at amounts found in 10% serum. Other molecules found in serum, specifically oleic acid and heparin, displace the ODN from the complex and thus interfere with delivery. This displacement is attenuated by first incubating the complex with BSA. Another manifestation of serum-complex interactions is that ODN significantly and cationic liposomes slightly, activate complement. However, formation of the complex markedly reduces the complement activation of the ODN. Finally, the effect of serum can be partially counteracted by the selection of the helper lipid (DOPE or cholesterol). Inclusion of a helper lipid reduces the effective charge ratio (cationic groups/anionic thioates) required to deliver ODN into cells and permits delivery in the presence of greater percentages of serum in the culture medium. These results support the current view that the binding of serum proteins to the complex is a significant factor in modulating the activity of cationic lipid-ODN complexes in culture and after intravenous administration. (C) 1998 Elsevier Science B.V.