Gene delivery to the rat retina by non-viral vectors based on chloroquine-containing cationic niosomes
Gene delivery to the rat retina by non-viral vectors based on chloroquine-containing cationic niosomes
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DOI：
10.1016/j.jconrel.2019.05.010
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发表时间：
2019-06-28
影响因子：
10.8
通讯作者：
Luis Pedraz, Jose
中科院分区：
文献类型：
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作者：
Mashal, Mohamed;Attia, Noha;Luis Pedraz, Jose
将氯喹掺入纳米制剂中，而不是作为细胞的共同治疗，可以为体内视网膜基因递送开辟一条新途径。在这篇手稿中，我们评估了二磷酸氯喹掺入由泊洛沙姆188、聚山梨酯80非离子表面活性剂和2，3-二（十四烷氧基）丙-1-胺（盐酸盐）阳离子脂质组成的阳离子囊泡制剂中，以抑制大鼠视网膜。通过反相蒸发技术制备不含和含二磷酸氯喹（分别为DPP 80和DPP 80-CQ）的囊泡制剂，并在尺寸、PDI、zeta电位和形态方面进行表征。在掺入pCMS-EGFP质粒后，进一步评价所得的纳米复合物（在不同的阳离子脂质/DNA质量比下）以压实、释放和保护DNA免受酶消化。在ARPE-19细胞中实现体外程序以评估转染功效和细胞内转运。两种nioplexes制剂都有效地转染了ARPE-19细胞，尽管细胞活力在DPP 80-CQ nioplexes的情况下明显更好。在视网膜下和玻璃体内注射后，DPP 80复合物不能抑制大鼠视网膜。然而，含有氯喹的载体在许多视网膜细胞中显示出蛋白质表达，具体取决于给药途径。这些数据为基于含氯喹的囊泡非病毒载体的视网膜基因递送提供了新的见解。
The incorporation of chloroquine within nano formulations, rather than as a co-treatment of the cells, could open a new avenue for in vivo retinal gene delivery. In this manuscript, we evaluated the incorporation of chloroquine diphosphate into the cationic niosome formulation composed of poloxamer 188, polysorbate 80 non-ionic surfactants, and 2,3-di (tetradecyloxy) propan-1-amine (hydrochloride salt) cationic lipid, to transfect rat retina. Niosome formulations without and with chloroquine diphosphate (DPP80, and DPP80-CQ, respectively) were prepared by the reverse phase evaporation technique and characterized in terms of size, PDI, zeta potential, and morphology. After the incorporation of the pCMS-EGFP plasmid, the resultant nioplexes -at different cationic lipid/DNA mass ratios-were further evaluated to compact, liberate, and secure the DNA against enzymatic digestion. In vitro procedures were achieved in ARPE-19 cells to assess transfection efficacy and intracellular transportation. Both nioplexes formulations transfected efficiently ARPE-19 cells, although the cell viability was clearly better in the case of DPP80-CQ nioplexes. After subretinal and intravitreal injections, DPP80 nioplexes were not able to transfect the rat retina. However, chloroquine containing vector showed protein expression in many retinal cells, depending on the administration route. These data provide new insights for retinal gene delivery based on chloroquine-containing niosome non-viral vectors.