Biodegradable nanoparticles decorated with different carbohydrates for efficient macrophage -targeted gene therapy
Biodegradable nanoparticles decorated with different carbohydrates for efficient macrophage -targeted gene therapy
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DOI：
10.1016/j.jconrel.2020.03.044
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发表时间：
2020-07-10
影响因子：
10.8
通讯作者：
Zhang, Xue-Qing
中科院分区：
文献类型：
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作者：
Chen, Qijing;Gao, Mingzhu;Zhang, Xue-Qing
由于它们对许多病理过程包括癌症、动脉粥样硬化、肥胖症、糖尿病和其他炎性疾病的贡献，大分子化合物是有吸引力的治疗靶点。巨噬细胞靶向基因治疗是调节炎症部位巨噬细胞功能治疗相关疾病的有效策略。然而，巨噬细胞被认为难以清除细胞，并且非特异性递送将不可避免地引起不希望的全身副作用。本文中，我们使用阳离子脂质样化合物G 0-C14和不同碳水化合物修饰的聚（丙交酯-共-乙交酯）（PLGA）或聚（丙交酯-共-乙交酯）-b-聚（乙二醇）（PLGA-PEG）通过稳健的自组装方法制备了一系列用于基因递送的巨噬细胞靶向纳米颗粒（NPs）。所产生的NP用基于碳水化合物的靶向部分（包括甘露糖、半乳糖、葡聚糖以及甘露糖和半乳糖的混合物）修饰。以EGFP信使RNA（mRNA）和GFP质粒DNA（pDNA）为报告基因，评价NP介导的基因转染巨噬细胞。巨噬细胞吞噬实验结果表明，更多的碳水化合物修饰的纳米粒子被Raw 264.7细胞比那些没有碳水化合物修饰的内吞。甘露糖修饰的纳米颗粒比仅用半乳糖修饰的纳米颗粒和甘露糖和半乳糖的混合物修饰的纳米颗粒显示出更好的对巨噬细胞的靶向能力。值得注意的是，多糖葡聚糖修饰的纳米粒也表现出明显的靶向作用。CCK-8测定显示，对于高达2.8 mg/mL的所有测试NP浓度，均未观察到细胞毒性。糖修饰的聚合物/G 0-C14对mRNA和pDNA表现出较强的包封作用，包封率超过95%。靶向的NP显著提高了巨噬细胞中的细胞内化和转染效率，这取决于NP表面上存在的碳水化合物部分的类型和含量。有趣的是，在巨噬细胞中在各种浓度下显示出更高内吞作用的葡聚糖修饰的NP也表现出更有效的mRNA转染，这表明NP介导的mRNA转染效率与内吞作用结果一致。
Macrophages are attractive therapeutic targets due to their contributions to many pathological processes including cancers, atherosclerosis, obesity, diabetes and other inflammatory diseases. Macrophage-targeted gene therapy is an effective strategy for regulating macrophage function at the site of inflammation to treat related diseases. However, macrophages are recognized as difficult to transfect cells and non-specific delivery would inevitably cause unwanted systemic side effects. Herein, we prepared a series of macrophage-targeted nanoparticles (NPs) using cationic lipid-like compound G0-C14 and different carbohydrates-modified poly(lactide-co-glycolide) (PLGA) or poly(lactide-coglycolide)-b-poly(ethylene glycol) (PLGA-PEG) for gene delivery by a robust self-assembly method. The yielded NPs were decorated with carbohydrate-based targeting moieties including mannose, galactose, dextran, and a mixture of mannose and galactose. EGFP messenger RNA (mRNA) and GFP plasmid DNA (pDNA) were used as reporter genes to evaluate NP-mediated gene transfection in macrophages. Experimental results of macrophage phagocytosis demonstrated that more carbohydrate-decorated NPs were endocytosed by Raw 264.7 cells than the ones without carbohydrate modification. Mannose-decorated NPs showed better targeting ability to macrophages than NPs decorated with galactose only and a blended mixture of mannose and galactose. It is worth noting that polysaccharide dextran-modified NPs also exhibited evident targeting effects. CCK-8 assay revealed that no cytotoxicity was observed for all tested NP concentrations up to 2.8 mg/mL. The carbohydrate-decorated polymer/G0-C14 exhibited strong entrapment of mRNA and pDNA with an encapsulation efficiency of above 95%. The targeted NPs significantly improved cellular internalization and transfection efficiency in macrophages, depending on the type and content of the carbohydrate moieties presented on the NP surface. Interestingly, dextran-decorated NPs showing higher endocytosis at various concentrations in macrophages also demonstrated more efficient mRNA transfection, suggesting that the NP-mediated mRNA transfection efficiency was consistent with the endocytosis results.