Transepithelial delivery of insulin conjugated with phospholipid-mimicking polymers via biomembrane fusion-mediated transcellular pathways
Transepithelial delivery of insulin conjugated with phospholipid-mimicking polymers via biomembrane fusion-mediated transcellular pathways
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通过生物膜融合介导的跨细胞途径，与磷脂模拟聚合物缀合的胰岛素经上皮递送
DOI：
10.1016/j.actbio.2021.12.003
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发表时间：
2022
影响因子：
9.7
通讯作者：
Goda Tatsuro
中科院分区：
文献类型：
--
作者：
Hatano Hiroaki;Meng Fanlu;Sakata Momoko;Matsumoto Akira;Ishihara Kazuhiko;Miyahara Yuji;Goda Tatsuro
上皮屏障通过形成紧密的连接来封闭细胞间隙，防止营养物质、电解质和药物的自由渗透，对于维持多细胞生物体的动态平衡是必不可少的。开发能够在不影响屏障功能的情况下渗透上皮组织的纳米载体是建立安全高效的药物传递系统(DDS)的关键。以前，我们已经证明了一种可溶于水的类磷脂随机共聚物，聚(2-甲基丙烯酰氧乙基磷酰胆碱30-随机正丁酯甲基丙烯酸酯70)(PMB30W)以被动扩散的方式进入活细胞的细胞质，而不破坏细胞膜。内化机制为两亲性诱导的膜融合。在本研究中，我们展示了PMB30W在体外通过Madin-Darby犬肾(MDCK)细胞单层模型上皮屏障的能量非依赖性渗透。该聚合物在不破坏屏障功能的情况下，穿透上皮MDCK单层和跨细胞途径。我们独特的检测证实了这一点，该检测可以监测质子的泄漏，作为跨越上皮屏障的最小指标。此外，当胰岛素与模拟磷脂的纳米载体化学偶联时，实现了能量不依赖的跨皮细胞渗透。胰岛素作为一种生长因子的生物活性被发现即使在易位后仍保持不变。这些基本发现可能有助于建立具有更高的药物效率和安全性的跨上皮DDS。意义陈述能够自由渗透上皮组织而不损害屏障功能的纳米载体是DDS成功的关键。现有的策略主要依赖于与紧密连接破坏相关的细胞旁运输或通过转运体识别介导的主动摄取的跨细胞运输。这些方法引发了人们对效率和安全的担忧。在这项研究中，我们在体外通过模型上皮屏障进行了磷脂模拟聚合物的非内吞渗透。聚合物通过跨细胞途径穿透，没有突破生物膜和紧密连接的屏障。此外，当胰岛素共价连接到纳米载体上时，会发生跨上皮渗透。即使在移位后，胰岛素的生物活性也保持不变。纳米载体的仿生设计可能实现安全、高效的跨上皮DDS。
Epithelial barriers that seal cell gaps by forming tight junctions to prevent the free permeation of nutrients, electrolytes, and drugs, are essential for maintaining homeostasis in multicellular organisms. The development of nanocarriers that can permeate epithelial tissues without compromising barrier function is key for establishing a safe and efficient drug delivery system (DDS). Previously, we have demonstrated that a water-soluble phospholipid-mimicking random copolymer, poly(2-methacryloyloxyethyl phosphorylcholine30-random-n‑butyl methacrylate70) (PMB30W), enters the cytoplasm of live cells by passive diffusion manners, without damaging the cell membranes. The internalization mechanism was confirmed to be amphiphilicity-induced membrane fusion. In the present study, we demonstrated energy-independent permeation of PMB30W through the model epithelial barriers of Madin-Darby canine kidney (MDCK) cell monolayersin vitro. The polymer penetrated epithelial MDCK monolayersviatranscellular pathways without breaching the barrier functions. This was confirmed by our unique assay that can monitor the leakage of the proton as the smallest indicator across the epithelial barriers. Moreover, energy-independent transepithelial permeation was achieved when insulin was chemically conjugated with the phospholipid-mimicking nanocarrier. The bioactivity of insulin as a growth factor was found to be maintained even after translocation. These fundamental findings may aid the establishment of transepithelial DDS with advanced drug efficiency and safety.Statement of significanceA nanocarrier that can freely permeate epithelial tissues without compromising barrier function is key for successful DDS. Existing strategies mainly rely on paracellular transport associated with tight junction breakdown or transcellular transport via transporter recognition-mediated active uptake. These approaches raise concerns about efficiency and safety. In this study, we performed non-endocytic permeation of phospholipid-mimicking polymers through the model epithelial barriersin vitro. The polymer penetrated via transcytotic pathways without breaching the barriers of biomembrane and tight junction. Moreover, transepithelial permeation occurred when insulin was covalently attached to the nanocarrier. The bioactivity of insulin was maintained even after translocation. The biomimetic design of nanocarrier may realize safe and efficient transepithelial DDS.