文献：反义寡脱氧核苷酸和小干扰RNA靶向递送至肺癌细胞

链接：https://pubs.acs.org/doi/abs/10.1021/mp060039w

作者：李世达，黄叶

摘要：

通过反义寡脱氧核苷酸 (AS-ODN) 或小干扰 RNA (siRNA) 疗法进行选择性基因抑制有望*传统药物无法治愈的疾病。然而，反义疗法由于在生理液中的稳定性差和细胞内摄取有限而受到阻碍。为了解决这些问题，我们实验室开发了一种配体靶向和空间稳定的纳米颗粒制剂。人类肺癌细胞通常过表达 σ 受体，因此可以用特定配体（例如茴香酰胺）进行靶向*。

将针对人类生存素的 AS-ODN 或 siRNA 与载体 DNA（小牛胸腺 DNA）混合，然后与鱼精蛋白（一种带高正电荷的肽）复合。将所得颗粒用由 DOTAP 和胆固醇（摩尔比 1:1）组成的阳离子脂质体包被，以获得 LPD（脂质体-聚阳离子-DNA）纳米颗粒。然后，通过后插入法将预制的LPD纳米粒子与DSPE-PEG-茴香酰胺（我们实验室早期开发的一种PEG化配体脂质）一起孵育，引入配体靶向性和空间稳定性。

同时制备了DSPE-PEG包覆的非靶向纳米粒子作为对照。通过Survivin mRNA下调、Survivin蛋白下调、引发*细胞凋亡的能力、*细胞生长抑制以及处理后的*细胞对*癌药物的化学增敏作用来测定纳米粒子的反义活性。我们发现，PEG化纳米粒子的*细胞递送和反义活性是序列依赖性的，并且依赖于茴香酰胺配体的存在。靶向PEG化纳米粒子对寡核苷酸的摄取可能会被过量的游离配体竞争。我们的结果表明，配体靶向和空间稳定的纳米粒子可以选择性地将 AS-ODN 和 siRNA 递送到肺癌细胞中进行*。

Abstract

Abstract Image

Selective gene inhibition by antisense oligodeoxynucleotide (AS-ODN) or by small interference RNA (siRNA) therapeutics promises the treatment of diseases that cannot be cured by conventional drugs. However, antisense therapy is hindered due to poor stability in physiological fluids and limited intracellular uptake. To address these problems, a ligand targeted and sterically stabilized nanoparticle formulation has been developed in our lab. Human lung cancer cells often overexpress the sigma receptor and, thus, can be targeted with a specific ligand such as anisamide. AS-ODN or siRNA against human survivin was mixed with a carrier DNA, calf thymus DNA, before complexing with protamine, a highly positively charged peptide. The resulting particles were coated with cationic liposomes consisting of DOTAP and cholesterol (1:1, molar ratio) to obtain LPD (liposome−polycation−DNA) nanoparticles. Ligand targeting and steric stabilization were then introduced by incubating preformed LPD nanoparticles with DSPE-PEG-anisamide, a PEGylated ligand lipid developed earlier in our lab, by the postinsertion method. Nontargeted nanoparticles coated with DSPE-PEG were also prepared as a control. Antisense activities of nanoparticles were determined by survivin mRNA down-regulation, survivin protein down-regulation, ability to trigger apoptosis in tumor cells, tumor cell growth inhibition, and chemosensitization of the treated tumor cells to anticancer drugs. We found that tumor cell delivery and antisense activity of PEGylated nanoparticles were sequence dependent and rely on the presence of anisamide ligand. The uptake of oligonucleotide in targeted, PEGylated nanoparticles could be competed by excess free ligand. Our results suggest that the ligand targeted and sterically stabilized nanoparticles can provide a selective delivery of AS-ODN and siRNA into lung cancer cells for therapy.

西安齐岳生物提供相关产品：

DSPE-PEG-CD19 mAb HD37（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-HD37Fab′（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-scFv(HD37-CCH)（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-mAb 2C5（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-Fab’of matuzumab or C225（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-Fab’of C225 mAb(cetuximab)（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-anti-EGFR scFv C10（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-F(ab’)2of GAH（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

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