A model for the analysis of nonviral gene therapy
A model for the analysis of nonviral gene therapy
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DOI：
10.1038/sj.gt.3302076
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发表时间：
2003-09-01
期刊：
影响因子：
5.1
通讯作者：
Giorgio, TD
中科院分区：
文献类型：
--
作者：
Banks, GA;Roselli, RJ;Giorgio, TD
需要进一步了解转基因在细胞和细胞内传递的机制，才能产生基因治疗的临床应用。在这项工作中设计的隔室和计算模型与以前的实验数据相结合，以定量地估计在体外转基因传递中跨越细胞屏障的质粒转移的速率常数。两个细胞研究之间的实验条件保持不变，只改变细胞类型，以研究不同细胞系之间的比率差异。对于主动传输和被动扩散，估计了每个势垒的两个速率常数。完整质粒跨细胞质和核屏障的移位率在不同细胞系之间存在差异。CV1细胞的速率(0.23h(1)胞浆，0.08h(-1)核)慢于HeLa细胞(1.87h(1)胞浆，0.45h(1)核)。通过比较每个屏障上完整的质粒转运率，核膜被鉴定为限速屏障。与HeLa细胞相比，CV1细胞中较慢的完整质粒转位与转基因效率的绝对能力降低有关。HeLa细胞生产绿色荧光蛋白的效率是CV1细胞的三倍，每一个完整的质粒被输送到细胞核。数学建模与实验研究相结合，可以提供对非病毒基因治疗的详细、定量的了解。
Further understanding of the mechanisms involved in cellular and intracellular delivery of transgene is needed to produce clinical applications of gene therapy. The compartmental and computational model designed in this work is integrated with data from previous experiments to quantitatively estimate rate constants of plasmid translocation across cellular barriers in transgene delivery in vitro. The experimental conditions between two cellular studies were held constant, varying only the cell type, to investigate how the rates differed between cell lines. Two rate constants were estimated per barrier for active transport and passive diffusion. Translocation rates of intact plasmid across the cytoplasmic and nuclear barriers varied between cell lines. CV1 cells were defined by slower rates (0.23 h(1) cytoplasmic, 0.08 h(-1) nuclear) than those of the HeLa cells (1.87 h(1) cytoplasmic, 0.45 h(1) nuclear). The nuclear envelope was identified as a rate-limiting barrier by comparing the rate of intact plasmid translocation at each barrier. Slower intact plasmid translocation in CV1 cells was correlated with a reduced absolute capacity for transgene efficiency in comparison with HeLa cells. HeLa cells were three times more efficient than CV1 cells at producing green fluorescent protein per intact plasmid delivered to the nucleus. Mathematical modeling coordinated with experimental studies can provide detailed, quantitative understanding of nonviral gene therapy.