基于磷酸胆碱的两性离子聚合物的 DNA 多聚物用于基因传递。
DNA Polyplexes of a Phosphorylcholine-Based Zwitterionic Polymer for Gene Delivery.
机构信息
Graduate School of Pharmaceutical Sciences, School of Pharmacy, Duquesne University, 600 Forbes Avenue, Pittsburgh, PA, 15282, USA.
Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37232, USA.
出版信息
Pharm Res. 2020 Aug 28;37(9):176. doi: 10.1007/s11095-020-02899-5.
PURPOSE
We tested polyplexes of a diblock polymer containing a pH-responsive, endosomolytic core (dimethylaminoethyl methacrylate and butyl methacrylate; DB) and a zwitterionic Poly (methacryloyloxyethyl phosphorylcholine) (PMPC) corona for the delivery of plasmid DNA (pDNA) to glioblastoma cells.
METHODS
We studied the physicochemical characteristics of the DNA polyplexes such as particle hydrodynamic diameter and surface potential. Cytocompatibility of free PMPC-DB polymer and pDNA polyplexes with U-87MG and U-138MG glioma cell lines were evaluated using the ATP assay. The transfection activity of luciferase pDNA polyplexes was measured using a standard luciferase assay. Anti-proliferative, apoptotic, and cell migration inhibitory activities of PMPC-DB/Interferon-beta (IFN-β1) pDNA polyplexes were examined using ATP assay, flow cytometry, and wound closure assay, respectively.
RESULTS
PMPC-DB copolymer condensed pDNA into nanosized polyplexes. DNA polyplexes showed particle diameters ranging from ca. 100-150 nm with narrow polydispersity indices and near electroneutral zeta potential values. PMPC-DB/Luciferase pDNA polyplexes were safe and showed an 18-fold increase in luciferase expression compared to the gold standard PEI polyplexes in U-87MG cells. PMPC-DB/IFN-β1 polyplexes induced apoptosis, demonstrated anti-proliferative effects, and retarded cell migration in glioblastoma cells.
CONCLUSION
The results described herein should guide the future optimization of PMPC-DB/DNA delivery systems for in vivo studies.
目的
我们测试了一种两亲嵌段共聚物的聚合物多聚物，该共聚物含有 pH 响应的内体溶解核心（二甲氨基乙基甲基丙烯酸酯和丁基甲基丙烯酸酯；DB）和两性离子聚（甲基丙烯酰氧基乙基磷酸胆碱）（PMPC）冠，用于将质粒 DNA（pDNA）递送至神经胶质瘤细胞。
方法
我们研究了 DNA 多聚物的物理化学特性，例如颗粒水动力直径和表面电位。使用 ATP 测定法评估游离 PMPC-DB 聚合物和 pDNA 多聚物与 U-87MG 和 U-138MG 神经胶质瘤细胞系的细胞相容性。使用标准的荧光素酶测定法测量荧光素酶 pDNA 多聚物的转染活性。使用 ATP 测定法、流式细胞术和划痕闭合测定法分别研究 PMPC-DB/干扰素-β（IFN-β1）pDNA 多聚物的抗增殖、凋亡和细胞迁移抑制活性。
结果
PMPC-DB 共聚物将 pDNA 凝聚成纳米级多聚物。DNA 多聚物的粒径在约 100-150nm 之间，具有较窄的多分散指数和接近电中性的 Zeta 电位值。PMPC-DB/Luciferase pDNA 多聚物是安全的，与金标准 PEI 多聚物相比，在 U-87MG 细胞中，荧光素酶表达增加了 18 倍。PMPC-DB/IFN-β1 多聚物诱导神经胶质瘤细胞凋亡，表现出抗增殖作用，并延缓细胞迁移。
结论
本文所述的结果应指导未来对 PMPC-DB/DNA 递药系统进行优化，以进行体内研究。
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