Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, United States.
Mol Pharm. 2012 May 7;9(5):1031-40. doi: 10.1021/mp200372s. Epub 2012 Apr 25.
This goal of this work was to explore histone H3 tail peptides containing transcriptionally activating modifications for their potential as gene delivery materials. We have found that these H3 tail peptides, in combination with the cationic polymer poly(ethylenimine) (PEI), can effectively bind and protect plasmid DNA. The H3/PEI hybrid polyplexes were found to transfect a substantially larger number of CHO-K1 cells in vitro compared to both polyplexes that were formed with only the H3 peptides and those that were formed with only PEI at the same total charge ratio; however, transfection was similarly high for polyplexes both with and without transcriptionally activating modifications. Transfections with the endolysosomal inhibitors chloroquine and bafilomycin A1 indicated that the H3/PEI hybrid polyplexes exhibited slower uptake and a reduced dependence on endocytic pathways that trafficked to the lysosome, indicating a potentially enhanced reliance on caveolar uptake for efficient gene transfer. In addition, whereas PEI polyplexes typically exhibit a cytotoxic effect, the H3/PEI hybrid polyplexes did not compromise cell viability. In total, the current studies provide new evidence for the potential role for histone-based materials as effective gene transfer agents, and support for the importance of subcellular trafficking for nonviral gene delivery.
这项工作的目的是探索含有转录激活修饰的组蛋白 H3 尾部肽作为基因传递材料的潜力。我们发现，这些 H3 尾部肽与阳离子聚合物聚（亚乙基亚胺）（PEI）结合，可以有效地结合并保护质粒 DNA。与仅由 H3 肽形成的和仅由相同总电荷比的 PEI 形成的复合聚合物相比，H3/PEI 杂交复合聚合物在体外转染 CHO-K1 细胞的数量要多得多；然而，对于具有和不具有转录激活修饰的复合聚合物，转染效率都很高。用内溶酶体抑制剂氯喹和巴弗洛霉素 A1 进行转染表明，H3/PEI 杂交复合聚合物的摄取速度较慢，对向溶酶体运输的内吞途径的依赖性降低，表明其对高效基因转移的 caveolar 摄取的潜在依赖性增强。此外，尽管 PEI 复合聚合物通常表现出细胞毒性作用，但 H3/PEI 杂交复合聚合物不会损害细胞活力。总的来说，目前的研究为基于组蛋白的材料作为有效的基因传递剂的潜在作用提供了新的证据，并支持了非病毒基因传递中细胞内运输的重要性。
