The elucidation of gene transferring mechanism by ultrasound-responsive unmodified and mannose-modified lipoplexes.
The elucidation of gene transferring mechanism by ultrasound-responsive unmodified and mannose-modified lipoplexes.
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DOI：
10.1016/j.biomaterials.2011.03.013
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发表时间：
2011-07
期刊：
影响因子：
14
通讯作者：
K. Un;S. Kawakami;Mitsuru Yoshida;Y. Higuchi;R. Suzuki;K. Maruyama;F. Yamashita;M. Hashida
中科院分区：
文献类型：
--
作者：
K. Un;S. Kawakami;Mitsuru Yoshida;Y. Higuchi;R. Suzuki;K. Maruyama;F. Yamashita;M. Hashida
开发简单、低毒条件下提高基因表达水平的基因转染方法是临床基因治疗的需要。本小组开发了超声（US）介导的甘露糖修饰基因载体man - peg2000泡脂丛基因转染方法，通过体外和体内暴露于US的man - peg2000泡脂丛基因转移，成功地获得了基因在甘露糖受体表达细胞中的选择性增强表达。在这里，我们研究了暴露于未修饰和man - peg2000气泡脂质体后的pDNA转移机制，特别关注了暴露时间。通过对细胞内转移特性的研究，我们将大量pDNA转移到细胞质中，然后使用US介导的气泡脂丛破坏气泡脂丛和US暴露的基因转移。此外，在加入泡状脂丛后，在不产生TNF-α的情况下，将US暴露至5分钟，可获得有效的基因表达。这些发现表明，使用未经修饰的和man - peg2000泡脂质体与US暴露进行基因转移可以将pDNA转移到细胞质中，优化US暴露时间对于实现高水平的基因表达和低水平的促炎细胞因子产生至关重要。
The development of gene transfection methods enhancing the level of gene expression under simple and low-toxic condition is required for gene therapy in clinical. Our group has developed the ultrasound (US)-mediated gene transfection method using Man-PEG2000bubble lipoplexes, which are US-responsive and mannose-modified gene carriers, and succeeded in obtaining the enhanced gene expression in mannose receptor-expressing cells selectively by the gene transfer using Man-PEG2000bubble lipoplexes with US exposurein vitroandin vivo. Here, we investigated pDNA transferring mechanism followed by US exposure to unmodified and Man-PEG2000bubble lipoplexes, in particular, focused on US exposure timing. Following investigation of intracellular transferring characteristics, a large amount of pDNA was transferred into the cytoplasm followed by US-mediated destruction of bubble lipoplexes in the gene transfer using both bubble lipoplexes with US exposure. Moreover, the effective gene expression was obtained without TNF-α production when US was exposed until 5 min after the addition of bubble lipoplexes. These findings suggest that the gene transfer using unmodified and Man-PEG2000bubble lipoplexes with US exposure enables to transfer pDNA into the cytoplasm, and optimized US exposure timing is important to achieve the high level of gene expression and the low level of pro-inflammatory cytokine production.