Optimization and Characterization of Protein Nanoparticles for the Targeted and Smart Delivery of Cytochrome c to Non-Small Cell Lung Carcinoma
Optimization and Characterization of Protein Nanoparticles for the Targeted and Smart Delivery of Cytochrome c to Non-Small Cell Lung Carcinoma
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DOI：
10.3390/cancers12051215
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发表时间：
2020-05-01
期刊：
影响因子：
5.2
通讯作者：
Ferrer-Acosta, Yancy
中科院分区：
文献类型：
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作者：
Barcelo-Bovea, Vanessa;Dominguez-Martinez, Irivette;Ferrer-Acosta, Yancy
细胞色素c（Cyt c）递送至胞质溶胶刺激细胞中的凋亡，其中其从线粒体的释放和凋亡诱导被抑制。我们开发了一种由叶酸-聚乙二醇-聚乳酸-羟基乙酸-硫醇（FA-PEG-PLGA-SH）修饰的Cyt c纳米颗粒组成的药物递送系统，以将Cyt c递送到癌细胞中，并在刘易斯肺癌（LLC）小鼠模型中测试其靶向。通过Cyt c纳米沉淀，随后用共聚物SH-PLGA-PEG-FA进行表面修饰，获得253 +/- 55和354 +/- 11 nm的Cyt c-PLGA-PEG-FA纳米颗粒（NP）。通过共聚焦显微镜证实Cytc-PLGA-PEG-FA纳米颗粒（NP）在LLC细胞中的内化。NP胱天蛋白酶活化比无NP制剂更有效。半胱天冬酶活性测定显示，NPs保留88-96%的Cyt c活性。NP制剂在降低LLC细胞活力方面比无NP制剂更有效，IC 50分别为49.2至70.1 μ g/mL和129.5 μ g/mL。我们的NP系统被证明对癌细胞的选择性是正常细胞的三倍。使用近红外标记的纳米颗粒的体内研究显示，注射后5分钟在小鼠LLC肿瘤中积累。总之，我们的细胞色素c的NP递送系统显示出优于无NP制剂的优势，并在免疫活性动物模型中达到叶酸过表达肿瘤。
The delivery of Cytochrome c (Cyt c) to the cytosol stimulates apoptosis in cells where its release from mitochondria and apoptotic induction is inhibited. We developed a drug delivery system consisting of Cyt c nanoparticles decorated with folate-poly(ethylene glycol)-poly(lactic-co-glycolic acid)-thiol (FA-PEG-PLGA-SH) to deliver Cyt c into cancer cells and tested their targeting in the Lewis Lung Carcinoma (LLC) mouse model. Cyt c-PLGA-PEG-FA nanoparticles (NPs) of 253 +/- 55 and 354 +/- 11 nm were obtained by Cyt c nanoprecipitation, followed by surface decoration with the co-polymer SH-PLGA-PEG-FA. The internalization of Cyt c-PLGA-PEG-FA nanoparticles (NPs) in LLC cells was confirmed by confocal microscopy. NP caspase activation was more efficient than the NP-free formulation. Caspase activity assays showed NPs retained 88-96% Cyt c activity. The NP formulations were more effective in decreasing LLC cell viability than NP-free formulation, with IC50 49.2 to 70.1 mu g/mL versus 129.5 mu g/mL, respectively. Our NP system proved to be thrice as selective towards cancerous than normal cells. In vivo studies using near infrared-tagged nanoparticles show accumulation in mouse LLC tumor 5 min post-injection. In conclusion, our NP delivery system for Cyt c shows superiority over the NP-free formulation and reaches a folic acid-overexpressing tumor in an immune-competent animal model.