Rapid crossing of the pulmonary endothelial barrier by polyethylenimine/DNA complexes
Rapid crossing of the pulmonary endothelial barrier by polyethylenimine/DNA complexes
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DOI：
10.1038/sj.gt.3301113
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发表时间：
2000-03-01
期刊：
影响因子：
5.1
通讯作者：
Demeneix, BA
中科院分区：
文献类型：
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作者：
Goula, D;Becker, N;Demeneix, BA
静脉给药可以成为预防性和治疗性基因治疗的一种递送途径，前提是基因能够穿过毛细血管屏障到达靶组织而不被降解。我们研究了线性聚乙烯亚胺（L-PEI）与DNA络合和静脉注射后转基因通过小鼠肺毛细血管的动力学和过程。使用地高辛标记的DNA，我们在注射后的不同时间跟踪DNA的细胞定位，并将这些发现与细胞标记和转基因表达相关联。注射后2小时，一些DNA仍定位在毛细血管管腔的下方，但其他复合物已经越过屏障，导致基因表达。注射后24小时，大部分标记的DNA定位在肺细胞中，转基因表达也是如此。只有很少在内皮细胞中发现转基因表达，这表明复合物可以迅速穿过毛细血管屏障。caspase-1样活性水平在转染后没有增加，这意味着L-PEI/DNA复合物通过非破坏性的生理过程通过细胞屏障运输，而不会引起炎症。不同转基因在肺细胞中的高水平表达表明，L-PEI/DNA复合物通过内皮屏障的运输不影响其转染能力。这些发现为基因传递及其在肺部的应用开辟了新的可能性。
Intravenous administration could become a delivery route of choice for prophylactic and curative gene therapies on condition that genes cross the capillary barrier and reach target tissues without being degraded. We investigated the kinetics and process of transgene delivery through mouse lung capillaries following DNA complexation with linear polyethylenimine (L-PEI) and intravenous injection. Using digoxin-labeled DNA we followed the cellular localization of DNA at different times after injection and correlated these findings with cell markers and transgene expression. At 2 h after injection some DNA was still localized on the inferior of the capillary lumen, but other complexes had already crossed the barrier and resulted in gene expression. At 24 h after injection most labeled DNA was localised in pulmonary cells, as was transgene expression. Only rarely was transgene expression found in endothelial cells, suggesting that the complexes cross the capillary barrier rapidly. Levels of caspase-1-like activity did not increase following transfection implying that L-PEI/DNA complexes are transported across cellular barriers by a non-damaging, physiological process, without causing inflammation. The high levels of expression of different transgenes in pneumocytes indicates that transport of L-PEI/DNA complexes through the endothelial barrier does not affect their transfection capacity. These findings open up new possibilities for gene delivery and ifs application to the lung.