新型可还原线性 L-赖氨酸修饰共聚物作为高效非病毒载体。

Novel reducible linear L-lysine-modified copolymers as efficient nonviral vectors.

机构信息

Department of Pharmacological and Pharmaceutical Sciences, 1441 Moursund Street, University of Houston, Texas Medical Center Campus, Houston, Texas 77030, USA.

出版信息

J Control Release. 2010 May 10;143(3):326-34. doi: 10.1016/j.jconrel.2009.12.032. Epub 2010 Jan 11.

DOI:10.1016/j.jconrel.2009.12.032

PMID:20067812

Abstract

The development of biodegradable gene delivery systems with high transfection efficiencies is paramount to the clinical translation of nonviral gene carriers. Therefore, to produce a biocompatible, reducible, effective and non-toxic gene delivery system, we have designed and synthesized novel reducible linear L-lysine modified copolymers (LLCs) as an alternative to high molecular weight poly(L-lysine) (PLL). The molecular weight (MW) of the copolymers was found to be approximately 3.2kDa with a polydispersity index of approximately 1.2. Gel retardation assays showed complete condensation of DNA at N/P ratios greater than 20/1 and exceptional LLC/pDNA polyplex stability during incubation with DNase I. Release of DNA from the polyplexes only occurred in the presence of the reducing agent dithiothreitol (DTT). The particle sizes of LLC/pDNA polyplexes were found to be between 100 and 231 nm with surface charges of 0.8-17 mV respectively. The transfection efficiencies of the polyplexes as determined with a luciferase assay showed that LLC polyplexes produced five times higher transfection efficiencies in HDF cells, three times higher transfection efficiencies in MCF-7 cells, and four times higher transfection efficiencies in MA cells as compared to the optimal PLL control. The LLC/pDNA polyplexes showed significantly lower cytotoxicities as compared to the control in HDF, MCF-7 and MA cells at certain N/P ratios. Therefore, these results suggest that these novel LLCs are efficient, reducible and biocompatible polymers for nonviral gene delivery.

摘要

可生物降解的基因传递系统的发展具有高转染效率对于非病毒基因载体的临床转化至关重要。因此，为了生产具有生物相容性、可还原、有效和无毒的基因传递系统，我们设计并合成了新型的还原线性 L-赖氨酸修饰共聚物（LLC）作为高分子量聚 L-赖氨酸（PLL）的替代品。共聚物的分子量（MW）约为 3.2kDa，多分散指数约为 1.2。凝胶阻滞实验表明，在 N/P 比大于 20/1 时，DNA 完全被凝聚，并且 LLC/pDNA 超螺旋体在与 DNA 酶 I 孵育时具有出色的稳定性。只有在还原剂二硫苏糖醇（DTT）存在的情况下，DNA 才从多聚物中释放出来。LLC/pDNA 超螺旋体的粒径在 100nm 至 231nm 之间，表面电荷分别为 0.8mV 至 17mV。用荧光素酶测定法测定的转染效率表明，与最佳 PLL 对照相比，LLC 多聚物在 HDF 细胞中产生了五倍高的转染效率，在 MCF-7 细胞中产生了三倍高的转染效率，在 MA 细胞中产生了四倍高的转染效率。与对照相比，在一定的 N/P 比下，LLC/pDNA 超螺旋体在 HDF、MCF-7 和 MA 细胞中的细胞毒性显著降低。因此，这些结果表明这些新型 LLC 是用于非病毒基因传递的有效、可还原和生物相容的聚合物。