Milk exosomes anchored with hydrophilic and zwitterionic motifs enhance mucus permeability for applications in oral gene delivery
Milk exosomes anchored with hydrophilic and zwitterionic motifs enhance mucus permeability for applications in oral gene delivery
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锚定有亲水性和两性离子基序的乳外泌体可增强粘液通透性，用于口腔基因递送应用
DOI：
10.1039/d3bm01089a
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发表时间：
2024
影响因子：
6.6
通讯作者：
Bajpayee, Ambika G.
中科院分区：
文献类型：
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作者：
Zhang, Chenzhen;Zhang, Hengli;Millán Cotto, Héctor A.;Boyer, Timothy L.;Warren, Matthew R.;Wang, Chia-Ming;Luchan, Joshua;Dhal, Pradeep K.;Carrier, Rebecca L.;Bajpayee, Ambika G.
外泌体由于其生物相容性和非免疫原性而成为递送药物和遗传物质的有前途的工具。然而，由于它们在苛刻的胃肠道（GI）环境中易降解并且穿过粘液-上皮屏障的运输受阻，因此在实现成功的口服递送方面存在挑战。为了克服这些挑战，我们开发了高纯度的牛乳外泌体（mExo）作为一种可扩展的有效的口服药物递送系统，它可以通过在其表面上掺入亲水性和两性离子基序来定制。在我们的研究中，我们观察到在引入亲水性和两性离子表面修饰后，天然猪肠粘液中的转运速率显著提高了2.5-4.5倍，如transwell设置和光漂白后荧光恢复（FRAP）分析所示。值得注意的是，由嵌段肽（BP）功能化的mExo，由在两端以嵌段排列的阳离子和阴离子氨基酸组成，在酸性胃环境中表现出上级耐受性（蛋白质回收率为84.8 ± 7.7%），并且表现出肠上皮细胞摄取增加2.5倍。此外，mExo和mExo-BP都证明了功能性siRNA的成功细胞内递送，导致在低剂量siRNA（5 pmol）下靶绿色荧光蛋白（GFP）基因表达的高达65%的抑制，而不引起显著的毒性。这些发现突出了用亲水性和两性离子基序修饰mExo用于有效口服递送siRNA疗法的巨大潜力。
Exosomes have emerged as a promising tool for the delivery of drugs and genetic materials, owing to their biocompatibility and non-immunogenic nature. However, challenges persist in achieving successful oral delivery due to their susceptibility to degradation in the harsh gastrointestinal (GI) environment and impeded transport across the mucus-epithelium barrier. To overcome these challenges, we have developed high-purity bovine milk exosomes (mExo) as a scalable and efficient oral drug delivery system, which can be customized by incorporating hydrophilic and zwitterionic motifs on their surface. In our study, we observed significantly improved transport rates by 2.5–4.5-fold in native porcine intestinal mucus after the introduction of hydrophilic and zwitterionic surface modifications, as demonstrated by transwell setup and fluorescence recovery after photobleaching (FRAP) analysis. Remarkably, mExo functionalized by a block peptide (BP), consisting of cationic and anionic amino acids arranged in blocks at the two ends, demonstrated superior tolerability in the acidic gastric environment (with a protein recovery rate of 84.8 ± 7.7%) and exhibited a 2.5-fold increase in uptake by intestinal epithelial cells. Furthermore, both mExo and mExo-BP demonstrated successful intracellular delivery of functional siRNA, resulting in up to 65% suppression of the target green fluorescence protein (GFP) gene expression at a low dose of siRNA (5 pmol) without causing significant toxicity. These findings highlight the immense potential of modifying mExo with hydrophilic and zwitterionic motifs for effective oral delivery of siRNA therapies.