Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 120-749, Republic of Korea.
Biomaterials. 2011 Nov;32(33):8654-62. doi: 10.1016/j.biomaterials.2011.07.075. Epub 2011 Aug 15.
Gene therapy technology is a powerful tool to elucidate the molecular cues that precisely regulate stem cell fates, but developing safe vehicles or mechanisms that are capable of delivering genes to stem cells with high efficiency remains a challenge. In this study, we developed a magnetically guided adeno-associated virus (AAV) delivery system for gene delivery to human neural stem cells (hNSCs). Magnetically guided AAV delivery resulted in rapid accumulation of vectors on target cells followed by forced penetration of the vectors across the plasma membrane, ultimately leading to fast and efficient cellular transduction. To combine AAV vectors with the magnetically guided delivery, AAV was genetically modified to display hexa-histidine (6xHis) on the physically exposed loop of the AAV2 capsid (6xHis AAV), which interacted with nickel ions chelated on NTA-biotin conjugated to streptavidin-coated superparamagnetic iron oxide nanoparticles (NiStNPs). NiStNP-mediated 6xHis AAV delivery under magnetic fields led to significantly enhanced cellular transduction in a non-permissive cell type (i.e., hNSCs). In addition, this delivery method reduced the viral exposure times required to induce a high level of transduction by as much as to 2-10 min of hNSC infection, thus demonstrating the great potential of magnetically guided AAV delivery for numerous gene therapy and stem cell applications.
基因治疗技术是阐明精确调控干细胞命运的分子线索的有力工具，但开发能够高效将基因递送至干细胞的安全载体或机制仍然是一个挑战。在这项研究中，我们开发了一种磁引导腺相关病毒（AAV）传递系统，用于将基因递送至人神经干细胞（hNSC）。磁引导 AAV 传递导致载体在靶细胞上迅速聚集，随后载体强行穿透质膜，最终导致快速高效的细胞转导。为了将 AAV 载体与磁引导传递相结合，对 AAV 进行了基因修饰，使其在 AAV2 衣壳的物理暴露环上显示六组氨酸（6xHis）（6xHis AAV），与镍离子螯合在 NTA-生物素偶联到链霉亲和素包被的超顺磁性氧化铁纳米颗粒（NiStNPs）上。在磁场下，NiStNP 介导的 6xHis AAV 传递导致非允许细胞类型（即 hNSC）中的细胞转导显著增强。此外，这种传递方法减少了所需的病毒暴露时间，将 hNSC 感染时所需的高转导水平所需的病毒暴露时间减少了多达 2-10 分钟，从而证明了磁引导 AAV 传递在众多基因治疗和干细胞应用中具有巨大潜力。
