Neuron-specific delivery of nucleic acids mediated by Tet1-modified poly(ethylenimine)
Neuron-specific delivery of nucleic acids mediated by Tet1-modified poly(ethylenimine)
复制标题
DOI：
10.1002/jgm.1062
复制
发表时间：
2007-08-01
影响因子：
3.5
通讯作者：
Pun, Suzie H.
中科院分区：
文献类型：
--
作者：
Park, In-Kyu;Lasiene, Jurate;Pun, Suzie H.
研究背景：开发用于中枢神经系统（CNS）的微创、非病毒基因载体是神经系统疾病分子治疗发展的重要技术目标。一种方法是将被运动神经元识别并逆行转运到CNS的材料外周递送。Tet 1是Boulis及其同事鉴定的具有破伤风毒素结合特征的肽，其特异性地与运动神经元相互作用并经历快速、逆行递送至细胞索马。方法合成Tet 1-PEI（Tet 1-poly（ethylenimine），Tet 1-PEI）和NT-PEI（neurotensin-PEI，NT-PEI），并与质粒DNA复合形成聚合物。通过流式细胞术和共聚焦显微镜评估复合物在分化的神经元样PC-12细胞中的结合和摄取。为了确定基因递送效率，将复合物暴露于分化的各个阶段的PC-12细胞。结果Tet 1-PEI和NT-PEI复合物与分化的PC-12细胞特异性结合。通过递送至非神经元细胞和通过与游离配体的竞争研究证实了相互作用的特异性。Tet 1-PEI复合物优先转染经历NGF诱导分化的PC-12细胞。最后，Tet 1-PEI复合物的神经元特异性结合在原代neurons.Conclusions中得到证实，这些研究表明Tet 1-PEI作为非侵入性CNS递送的神经元靶向材料的潜力。Tet 1-PEI特异性结合并被神经元样PC-12细胞和初级背根神经节内化。未来的工作将包括这些载体的siRNA交付的评价。版权所有（c）2007约翰威利父子有限公司。
Background The development of minimally invasive, non-viral gene delivery vehicles for the central nervous system (CNS) is an important technology goal in the advancement of molecular therapies for neurological diseases. One approach is to deliver materials peripherally that are recognized and retrogradely transported by motor neurons toward the CNS. Tet1 is a peptide identified by Boulis and coworkers to possess the binding characteristics of tetanus toxin, which interacts specifically with motor neurons and undergoes fast, retrograde delivery to cell soma. In this work, Tet1-poly(ethylenimine) (Tet1-PEI) was synthesized and evaluated as a neurontargeted delivery vehicle.Methods Tet1-PEI and NT-PEI (neurotensin-PEI) were synthesized and complexed with plasmid DNA to form polyplexes. Polyplexes were assessed for binding and uptake in differentiated neuron-like PC-12 cells by flow cytometry and confocal microscopy. In order to determine gene delivery efficiency, polyplexes were exposed to PC-12 cells at various stages of differentiation. Targeted binding of polyplexes with primary neurons was studied using dorsal root ganglion cells.Results Tet1-PEI and NT-PEI polyplexes bound specifically to differentiated PC-12 cells. The specificity of the interaction was confirmed by delivery to non-neuronal cells and by competition studies with free ligands. Tet1-PEI polyplexes preferentially transfected PC-12 cells undergoing NGF-induced differentiation. Finally, neuron-specific binding of Tet1-PEI polyplexes was confirmed in primary neurons.Conclusions These studies demonstrate the potential of Tet1-PEI as a neuron-targeted material for non-invasive CNS delivery. Tet1-PEI binds specifically and is internalized by neuron-like PC-12 cells and primary dorsal root ganglion. Future work will include evaluation of siRNA delivery with these vectors. Copyright (c) 2007 John Wiley & Sons, Ltd.