Differential uptake of DNA-poly(ethylenimine) polyplexes in cells cultured on collagen and fibronectin surfaces.
Differential uptake of DNA-poly(ethylenimine) polyplexes in cells cultured on collagen and fibronectin surfaces.
复制标题
DOI：
10.1016/j.actbio.2010.03.038
复制
发表时间：
2010-09
影响因子：
9.7
通讯作者：
Segura T
中科院分区：
文献类型：
--
作者：
Dhaliwal A;Maldonado M;Han Z;Segura T
基因修饰的骨髓间充质干细胞（MSCs）已被证明是局部或全身递送治疗药物以及增加组织形成的生长因子的有效细胞载体。然而，有效的非病毒基因转移到这些细胞限制了它们的适用性。虽然大多数研究都集中在设计更有效的DNA凝聚剂上，但我们在这篇论文中的重点是研究两种细胞外基质蛋白对间充质干细胞转染能力的作用，胶原I （Col I）和纤维连接蛋白（Fn）。在这里，我们报道了将MSCs镀在Col - i涂层表面抑制转染，而将MSCs镀在fn涂层表面则增强转染。这些ECM蛋白影响非病毒基因转移的机制涉及用于多重摄取和细胞内张力的内吞作用途径。我们发现Fn通过网格蛋白介导的内吞作用促进内化，并且这种途径比小泡介导的内吞作用和巨噬细胞作用更有效。此外，肌动蛋白-肌球蛋白相互作用的破坏导致镀在Fn涂层表面的细胞基因转移增强，而镀在Col i上的细胞则没有。我们相信细胞微环境可以被改造以增强细胞被转染的能力，并且通过了解ECM影响非病毒基因转移的机制，可以实现更好的材料和转染方案。
Genetically modified bone marrow derived mesenchymal stem cells (MSCs) have proven to be efficient cell carriers for local or systemic delivery of therapeutics as well as for growth factors to augment tissue formation. However, efficient non-viral gene transfer to these cells is limiting their applicability. Although most studies focus on designing more efficient condensation agents for DNA, our focus in this manuscript is to study the role of two extracellular matrix proteins on the ability of MSCs to become transfected, collagen I (Col I) and fibronectin (Fn). Here we report that plating MSCs on Col I-coated surfaces inhibits transfection, while plating MSCs on Fn-coated surfaces enhances transfection. The mechanism by which these ECM proteins affect non-viral gene transfer involves the endocytosis pathway used for polyplex uptake and intracellular tension. We found that Fn promoted internalization through clathrin-mediated endocytosis and that this pathway resulted in more efficient transfection than caveolae-mediated endocytosis and macropinocytosis. Further, the disruption of actin-myosin interactions resulted in an enhancement of gene transfer for cells plated on Fn coated surfaces, but not for cells plated on Col I. We believe that the cellular microenvironment can be engineered to enhance the ability of cells to become transfected and that through understanding the mechanisms by which the ECM affects non-viral gene transfer, better materials and transfection protocols can be realized.
登录
查看更多内容
影响因子：
3.7
作者：
Bhadriraju, Kiran;Yang, Michael;Chen, Christopher S.
通讯作者：
Chen, Christopher S.
影响因子：
10.8
作者：
Kong, Hyun Joon;Hsiong, Susan;Mooney, David J.
通讯作者：
Mooney, David J.
诺考达唑对脂质双层包被的金纳米粒子转染效率的影响。
影响因子：
14
作者：
Li, Dan;Li, Peicai;Wang, Erkang
通讯作者：
Wang, Erkang
影响因子：
41.2
作者：
Kong, HJ;Liu, JD;Mooney, DJ
通讯作者：
Mooney, DJ
影响因子：
3.5
作者：
Kopatz, I;Remy, JS;Behr, JP
通讯作者：
Behr, JP