A nanogel with passive targeting function and adjustable polyplex surface properties for efficient anti-tumor gene therapy
A nanogel with passive targeting function and adjustable polyplex surface properties for efficient anti-tumor gene therapy
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DOI：
10.1039/c6ra13707e
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发表时间：
2016-09
期刊：
影响因子：
3.9
通讯作者：
Hai-zhou Zhang;Qingbao Li;Yingying Zhang;Yu Xia;Li Yun;Qian Zhang;Tao Zhang;Xia Chen;Huaiwen Chen;Wei Li
中科院分区：
文献类型：
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作者：
Hai-zhou Zhang;Qingbao Li;Yingying Zhang;Yu Xia;Li Yun;Qian Zhang;Tao Zhang;Xia Chen;Huaiwen Chen;Wei Li
具有高转染效率和血清稳定性的非细胞毒性载体对于成功的基因递送起着关键作用，这在很大程度上取决于复合物的结构特性和细胞亲和力。在本研究中，通过用非细胞毒性单体N-异丙基丙烯酰胺（NIPAM）修饰“金标准”转染剂聚乙烯亚胺（PEI），我们成功地开发了温敏阳离子PNIPAM/PEI纳米凝胶。它具有交联的热敏核和阳离子壳，具有可调节的尺寸、表面电位和被动细胞靶向功能。采用激光光散射法对纳米凝胶的尺寸、表面电位、热敏性和血清稳定性等复合物性质进行了系统的研究。结果发现，通过纳米凝胶的体外转移效率比PEI高约两倍，由于被动细胞靶向，当T > VPTT时，其进一步提高约两倍。通过倒置荧光显微镜、流式细胞仪和激光共聚焦显微镜研究了纳米凝胶的体外细胞摄取、基因转移效率及相关机制。还在Balb/c裸小鼠异种移植肿瘤模型中评估了其生物分布和高瘤内积聚。这种复杂的纳米凝胶显著抑制肿瘤生长，体积比PEI小5倍，这表明其在实际基因治疗中的高效性。
Non-cytotoxic vectors with high transfection efficiency and serum stability play a key role for successful gene delivery, which is strongly determined by polyplex structural properties and cellular affinity. In this study, through modifying the “gold-standard” transfection agent poly(ethylenimine) (PEI) with non-cytotoxic monomer N-isopropylacrylamide (NIPAM), we successfully developed a thermosensitive cationic PNIPAM/PEI nanogel. It has a crosslinked thermosensitive core and cationic shell with adjustable dimensions, surface potential and passive cellular targeting function. The polyplex properties such as size, surface potential, thermo-sensitivity and serum stability of the nanogel was systemically investigated by laser light scattering. It was found that the in vitro transfer efficiency by the nanogel was about two times higher than that of PEI, which was further enhanced about two time as T > VPTT due to the passive cellular targeting. The obviously enhanced in vitro cellular uptake, gene transfer efficiency and corresponding mechanism of the nanogel were then revealed by inverse fluorescent microscopy, a flow cytometer and confocal laser scanning microscopy. Its biodistribution and high intratumor accumulation were also evaluated in a Balb/c nude mice xenograft tumor model. Such a sophisticated nanogel significantly suppressed tumor growth with a volume 5 times smaller than that of PEI, which indicated its high potent for practical gene therapy.