A model for non-viral gene delivery: through syndecan adhesion molecules and powered by actin
A model for non-viral gene delivery: through syndecan adhesion molecules and powered by actin
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DOI：
10.1002/jgm.558
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发表时间：
2004-07-01
影响因子：
3.5
通讯作者：
Behr, JP
中科院分区：
文献类型：
--
作者：
Kopatz, I;Remy, JS;Behr, JP
细胞转染需要阳离子DNA复合物和细胞表面的硫酸乙酰肝素蛋白聚糖（HSPG）。多配体蛋白聚糖是在贴壁细胞上广泛表达的跨膜HSPG。其聚阴离子硫酸乙酰肝素部分结合在其胞外域的远端，从而促进与大阳离子颗粒的相互作用。方法我们提出了一个细胞进入模型，涉及syndecans作为DNA复合物的受体，通过比较转染与细菌摄取，并使用药物抑制实验沿着与共聚焦显微镜。结果当结合文献的结果，我们的数据表明，以下顺序的事件：初始颗粒结合后，逐渐静电拉链的质膜上的颗粒是持续的syndecan分子簇成富含胆固醇的筏横向扩散。聚簇反过来触发PKC活性和连接蛋白介导的肌动蛋白结合到多配体蛋白聚糖的胞质尾部。由此产生的张力纤维和不断增长的皮层肌动蛋白网络可以将颗粒拉入细胞。结论转移整合素和syndecan介导的细胞粘附过程颗粒吞噬似乎被广泛利用的动物（乳糜微粒），病原体（细菌，病毒），并在这里建议，由非病毒载体。版权所有（C）2004约翰威利父子有限公司。
Background Cell transfection requires cationic DNA complexes and heparan sulfate proteoglycans (HSPGs) at the cell surface. Syndecans are transmembrane HSPGs that are ubiquitously expressed on adherent cells. Their polyanionic heparan sulfate moieties are bound at the distal end of their ectodomain, thus facilitating interaction with large cationic particles. Methods We propose a model for cell entry involving syndecans as receptors for the DNA complexes by comparing transfection with bacteria uptake and using drug inhibition experiments along with confocal microscopy. Results When combined with results from the literature, our data suggest the following sequence of events: after initial particle binding, gradual electrostatic zippering of the plasma membrane onto the particle is sustained by lateral diffusion of syndecan molecules that cluster into cholesterol-rich rafts. Clustering in turn triggers PKC activity and linker protein-mediated actin binding to the cytoplasmic tail of the syndecans. Resulting tension fibers and a growing network of cortical actin may then pull the particle into the cell. Conclusions Diversion of integrin- and syndecan-mediated cell adhesion processes for particle engulfment appears to be widely exploited by animals (chylomicrons), by pathogens (bacteria, viruses) and, as suggested here, by non-viral vectors. Copyright (C) 2004 John Wiley Sons, Ltd.