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N-异丙基丙烯酰胺与甲基丙烯酸2-二甲基（氨基乙基）酯共聚物在用于鼻咽癌基因递送的非病毒和腺病毒载体中的评估

Evaluation of copolymers of N-isopropylacrylamide and 2-dimethyl(aminoethyl)methacrylate in nonviral and adenoviral vectors for gene delivery to nasopharyngeal carcinoma.

作者信息

Moselhy Jim, Sarkar Swapna, Chia Maria C, Mocanu Joseph D, Taulier Nicolas, Liu Fei-Fei, Wu Xiao Yu

机构信息

Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Canada.

出版信息

Int J Nanomedicine. 2007;2(3):461-78.

PMID:18019844
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2676666/
Abstract

Copolymers of 2-dimethyl(aminoethyl) methacrylate (PDMAEM) with N-isopropylacrylamide (NIPAM) were evaluated for their potential to enhance transgene expression of plasmid DNA (pDNA) and gene delivery by adenovirus vectors. The polymers of varying compositions and molecular weights (MW) were synthesized by free-radical polymerization. Polyelectrolyte complexes (PECs) were prepared with different charge (N:P) ratios of PNIPAM/ DMAEM to pDNA. Polymer-modified viral vectors based on non-replicating adenovirus serotype 5 (Ad5), (deltaE1/oriP/luc) or (deltaE1/CMV/luc) transcriptor/promoter/reporter were constructed by electrostatically coupling PNIPAM/DMAEM (Type 1) or PECs (oriP/luc, 6.6 kb) (Type II) to the viral capsid. The N:P value at complete condensation was lower for PECs with higher DMAEM content and MW. pDNA binding was enhanced by high MW PNIPAM/DMAEM. Circular dichroism spectroscopy revealed changes to the secondary structure of pDNA and adenovirus capsid proteins in the presence of PNIPAM/DMAEM. The toxicity of PNIPAM/DMAEM to CNE-1 nasopharyngeal cancer (NPC) cells diminished with decreasing DMAEM content and increasing MW The transfection efficiency ofC666-1 NPC cells by PECs increased with DMAEM content and MW. The transduction efficiency of CNE-1 NPC cells by Type I Ad5 vectors improved with DMAEM content, but was independent of MW. The transduction efficiency of Type II Ad5 in C666-1 cells approximated the sum of expression levels of the PECs and Ad5 vectors individually. PDMAEM and PNIPAM/DMAEM demonstrate both transfection and transduction enhancement activity of modified vectors in nasopharyngeal cancer cells in culture.

摘要

评估了甲基丙烯酸2 - 二甲基（氨基乙基）酯（PDMAEM）与N - 异丙基丙烯酰胺（NIPAM）的共聚物增强质粒DNA（pDNA）转基因表达以及腺病毒载体基因递送的潜力。通过自由基聚合合成了不同组成和分子量（MW）的聚合物。用不同电荷（N:P）比的PNIPAM/DMAEM与pDNA制备了聚电解质复合物（PEC）。基于非复制性腺病毒血清型5（Ad5）、（deltaE1/oriP/luc）或（deltaE1/CMV/luc）转录子/启动子/报告基因构建了聚合物修饰的病毒载体，通过将PNIPAM/DMAEM（I型）或PEC（oriP/luc，6.6 kb）（II型）静电偶联到病毒衣壳上。DMAEM含量和MW较高的PEC在完全凝聚时的N:P值较低。高分子量的PNIPAM/DMAEM增强了pDNA结合。圆二色光谱显示在PNIPAM/DMAEM存在下pDNA和腺病毒衣壳蛋白的二级结构发生了变化。PNIPAM/DMAEM对CNE - 1鼻咽癌（NPC）细胞的毒性随着DMAEM含量的降低和MW的增加而减弱。PEC对C666 - 1 NPC细胞的转染效率随着DMAEM含量和MW的增加而提高。I型Ad5载体对CNE - 1 NPC细胞的转导效率随着DMAEM含量的增加而提高，但与MW无关。II型Ad5在C666 - 1细胞中的转导效率近似于PEC和Ad5载体各自表达水平的总和。PDMAEM和PNIPAM/DMAEM在培养的鼻咽癌细胞中均表现出修饰载体的转染和转导增强活性。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/5caa874544d6/ijn-2-461f13.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/e372b090a56c/ijn-2-461f1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/a4ad2faa6924/ijn-2-461f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/bad170725332/ijn-2-461f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/125dec0c4a32/ijn-2-461f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/b9b24318bb0c/ijn-2-461f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/a98a06a6fe1a/ijn-2-461f9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/59e5347b0294/ijn-2-461f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/e60faadfbf99/ijn-2-461f11.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/74e2ab628d2a/ijn-2-461f12.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c96e/2676666/5caa874544d6/ijn-2-461f13.jpg

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