Plasmid-loadable magnetic/ultrasound-responsive nanodroplets with a SPIO-NP dispersed perfluoropentane core and lipid shell for tumor-targeted intracellular plasmid delivery
Plasmid-loadable magnetic/ultrasound-responsive nanodroplets with a SPIO-NP dispersed perfluoropentane core and lipid shell for tumor-targeted intracellular plasmid delivery
复制标题
具有 SPIO-NP 分散全氟戊烷核和脂质壳的可负载质粒的磁性/超声响应纳米滴，用于肿瘤靶向细胞内质粒递送
DOI：
10.1039/d0bm00699h
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发表时间：
2020-10-07
影响因子：
6.6
通讯作者：
Zong, Yujin
中科院分区：
文献类型：
--
作者：
Dong, Wei;Huang, Anqi;Zong, Yujin
使用超声激活造影剂诱导声孔作用是有效靶向病变基因递送的潜在策略。此前的报道已经证明，亚微米纳米液滴比微泡具有更好的优势，因为它们可以通过被动靶向的方式穿过肿瘤脉管系统内皮间隙；然而，它们无法在肿瘤中达到足够的剂量来促进超声增强的基因传递。此外，一些专注于传递大分子遗传材料（即过表达质粒和CRISPR质粒）的研究显示出比小分子遗传材料（即miRNA模拟物、siRNA和shRNA等）更独特的优势，例如增强目标基因的表达并具有长期有效性。因此，我们构建了新型可负载质粒的磁性/超声响应纳米液滴，其中分散的全氟戊烷中的超顺磁性氧化铁纳米颗粒被脂质封装，质粒可以粘附在其上，并使用支链聚乙烯亚胺来保护质粒免于酶解。此外，还进行了体外和体内研究，以验证可负载质粒的磁性/超声响应纳米滴的磁性肿瘤靶向能力以及聚焦超声增强的细胞内质粒递送。可负载质粒的磁性/超声响应纳米液滴，每个液滴携带16-19个质粒，具有小于300 nm的理想直径，并结合了优异的磁性靶向能力和对聚焦超声的相变敏感性的优点。在可编程聚焦超声照射下，可装载质粒的磁性/超声响应纳米液滴经历相变，形成回声微泡，随后微泡的惯性空化实现了类似于 40% 的体外质粒递送效率。静脉内给药后，肿瘤的T2加权磁共振成像、扫描电子显微镜和电感耦合等离子体发射光谱显示，在外部磁场下，可负载质粒的磁性/超声响应纳米液滴的瘤内积累显着增强。 GFP ELISA 测定和免疫荧光染色表明，聚焦超声诱导的可装载质粒的磁性/超声响应纳米滴的惯性空化显着增强了磁体辅助积累后肿瘤内质粒的细胞内递送，而仅应用聚焦超声或单独的外部磁体时，在肿瘤中仅观察到较低的 GFP 水平。综上所述，利用优异的可负载质粒的磁性/超声响应纳米液滴，结合磁性和超声，可以有效地将质粒递送至癌细胞，这可能是临床上治疗癌症的大分子遗传物质递送的潜在策略。
Using ultrasound activating contrast agents to induce sonoporation is a potential strategy for effective lesion-targeted gene delivery. Previous reports have proven that submicron nanodroplets have a better advantage than microbubbles in that they can pass through tumor vasculature endothelial gaps by passive targeting; however, they cannot achieve an adequate dose in tumors to facilitate ultrasound-enhanced gene delivery. Additionally, a few studies focused on delivering macromolecular genetic materials (i.e.overexpression plasmid and CRISPR plasmid) have presented more unique advantages than small-molecular genetic materials (i.e.miRNA mimics, siRNA and shRNAetc.), such as enhancing the expression of target genes with long-term effectiveness. Thereby, we constructed novel plasmid-loadable magnetic/ultrasound-responsive nanodroplets, where superparamagnetic iron oxide nanoparticle dispersed perfluoropentane was encapsulated with lipids to which plasmids could be adhered, and branched polyethylenimine was used to protect the plasmids from enzymolysis. Furthermore,in vitroandin vivostudies were performed to verify the magnetic tumor-targeting ability of the plasmid-loadable magnetic/ultrasound-responsive nanodroplets and focused ultrasound enhanced intracellular plasmid delivery. The plasmid-loadable magnetic/ultrasound-responsive nanodroplets, carrying 16-19 plasmids per droplet, had desirable diameters less than 300 nm, and integrated the merits of excellent magnetic targeting capabilities and phase transition sensitivity to focused ultrasound. Under programmable focused ultrasound exposure, the plasmid-loadable magnetic/ultrasound-responsive nanodroplets underwent a phase-transition into echogenic microbubbles and the subsequent inertial cavitation of the microbubbles achieved an similar to 40%in vitroplasmid delivery efficiency. Following intravenous administration, T2-weighted magnet resonance imaging, scanning electron microscopy and inductively coupled plasma optical emission spectrometry of the tumors showed significantly enhanced intratumoral accumulation of the plasmid-loadable magnetic/ultrasound-responsive nanodroplets under an external magnetic field. And a GFP ELISA assay and immunofluorescence staining indicated that focused ultrasound-induced inertial cavitation of the plasmid-loadable magnetic/ultrasound-responsive nanodroplets significantly enhanced the intracellular delivery of plasmids within the tumor after magnet-assisted accumulation, while only lower GFP levels were observed in the tumors on applying focused ultrasound or an external magnet alone. Taken together, utilizing the excellent plasmid-loadable magnetic/ultrasound-responsive nanodroplets combined with magnetism and ultrasound could efficiently deliver plasmids to cancer cells, which could be a potential strategy for macromolecular genetic material delivery in the clinic to treat cancer.