Milk exosomes with enhanced mucus penetrability for oral delivery of siRNA.
Milk exosomes with enhanced mucus penetrability for oral delivery of siRNA.
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DOI：
10.1039/d0bm01497d
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发表时间：
2021-06-15
影响因子：
6.6
通讯作者：
Bajpayee AG
中科院分区：
文献类型：
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作者：
Warren MR;Zhang C;Vedadghavami A;Bokvist K;Dhal PK;Bajpayee AG
牛乳来源的外泌体最近成为一种有前途的纳米载体，用于封装和递送大分子生物治疗药物。在这里，我们设计了高纯度牛乳外泌体（mExo），具有模块化表面可调性，用于口服递送小干扰RNA（siRNA）。我们利用一种低成本的富集方法，结合酪蛋白螯合与微分超离心，然后通过尺寸排阻色谱法，产生高浓度和纯度的mExo。使用体外模型，我们证明了带负电荷的疏水mExos可以穿透多种生物屏障以进行口服药物递送。通过偶联脂质尾的被动、稳定的疏水插入在mExo表面上引入亲水性聚乙二醇（PEG）涂层，与未修饰的mExo相比，其显著降低了mExo在酸性胃环境中的降解，并将其通过粘蛋白的渗透性提高了3倍以上。mExo和PEG-mExo均表现出肠上皮细胞的高摄取，并介导siRNA的功能性细胞内递送，从而抑制靶绿色荧光蛋白（GFP）基因的表达高达70%。我们还表明，阳离子化学转染是显着更有效地加载siRNA到mExo比电穿孔。本文所展示的分离高浓度的高纯度mExo并使其具有可调表面性质的简单性为mExo作为用于siRNA口服药物递送的有效、可扩展的平台技术的开发铺平了道路。高纯度的牛奶外泌体被设计成具有表面PEG化和siRNA包封，用于增强口服药物递送。
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 that significantly reduced mExo degradation in acidic gastric environment and enhanced their permeability through mucin by over 3x 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. High-purity milk exosomes were engineered with surface PEGylation and siRNA encapsulation for enhanced oral drug delivery.