增强黏液穿透性的牛奶外泌体用于口服递送 siRNA。
Milk exosomes with enhanced mucus penetrability for oral delivery of siRNA.
机构信息
Departments of Bioengineering, Northeastern University, Boston, MA 02115, USA.
Sanofi SA Global R&D, Frankfurt am Main, Germany.
出版信息
Biomater Sci. 2021 Jun 15;9(12):4260-4277. doi: 10.1039/d0bm01497d.
Bovine milk-derived exosomes have recently emerged as a promising nano-vehicle for the encapsulation and delivery of macromolecular biotherapeutics. Here we engineer high purity bovine milk exosomes (mExo) with modular surface tunability for oral delivery of small interfering RNA (siRNA). We utilize a low-cost enrichment method combining casein chelation with differential ultracentrifugation followed by size exclusion chromatography, yielding mExo of high concentration and purity. Using in vitro models, we demonstrate that negatively charged hydrophobic mExos can penetrate multiple biological barriers to oral drug delivery. A hydrophilic polyethylene glycol (PEG) coating was introduced on the mExo surface via passive, stable hydrophobic insertion of a conjugated lipid tail, which significantly reduced mExo degradation in acidic gastric environment and enhanced their permeability through mucin by over 3× compared to unmodified mExo. Both mExo and PEG-mExo exhibited high uptake by intestinal epithelial cells and mediated functional intracellular delivery of siRNA, thereby suppressing the expression of the target green fluorescence protein (GFP) gene by up to 70%. We also show that cationic chemical transfection is significantly more efficient in loading siRNA into mExo than electroporation. The simplicity of isolating high purity mExo in high concentrations and equipping them with tunable surface properties, demonstrated here, paves way for the development of mExo as an effective, scalable platform technology for oral drug delivery of siRNA.
牛源外泌体最近作为一种有前途的纳米载体,用于封装和输送大分子生物治疗药物。在这里,我们通过模块化表面可调节性工程设计了高纯度牛源外泌体(mExo),用于口服递送小干扰 RNA(siRNA)。我们利用一种结合了酪蛋白螯合与差速超速离心的低成本富集方法,随后进行分子筛层析,得到高浓度和高纯度的 mExo。通过体外模型,我们证明了带负电荷的疏水 mExo 可以穿透多种生物屏障进行口服药物递送。通过被动、稳定的疏水插入共轭脂质尾巴,在 mExo 表面引入了亲水性聚乙二醇(PEG)涂层,与未修饰的 mExo 相比,这显著降低了 mExo 在酸性胃环境中的降解,并使其通过粘蛋白的通透性增强了 3 倍以上。mExo 和 PEG-mExo 均被肠道上皮细胞高效摄取,并介导 siRNA 的功能性细胞内递送,从而使靶绿色荧光蛋白(GFP)基因的表达抑制高达 70%。我们还表明,与电穿孔相比,阳离子化学转染在将 siRNA 加载到 mExo 中更为高效。这里展示的高纯度 mExo 的简单高效分离方法,以及对其表面性质的可调节性,为开发 mExo 作为有效、可扩展的 siRNA 口服药物递送平台技术铺平了道路。
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