Using a Membrane-Penetrating-Peptide to Anchor Ligands in the Liposome Membrane Facilitates Targeted Drug Delivery.
Using a Membrane-Penetrating-Peptide to Anchor Ligands in the Liposome Membrane Facilitates Targeted Drug Delivery.
复制标题
使用穿膜肽将配体锚定在脂质体膜上有利于靶向药物递送
DOI：
10.1021/acs.bioconjchem.9b00798
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发表时间：
2020
影响因子：
4.7
通讯作者：
Wu Guoqiu
中科院分区：
文献类型：
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作者：
Fan Xiaobo;Xu Hongbo;Song Junlong;Jin Yongcan;Wink Michael;Wu Guoqiu
抗菌肽（Antimicrobial peptides，AMP）是一类典型的细胞穿透肽（cellpenetrating peptides，CPPs），具有广泛的生物活性。我们设计了一个三重融合蛋白，由AMP、N端的Ib-AMP 4、中心的荧光GFP探针和另一端的肿瘤靶向肽P1 c组成。弗罗姆. coli中表达，研究了Ib-AMP 4-GFP-P1 c融合蛋白（IGP）与脂膜的相互作用。等温滴定量热法（ITC）和石英晶体微天平与耗散（QCM-D）的实验表明，IGP蛋白自发结合的脂质双层的最大摩尔比为1：52（蛋白质：脂质）。此外，透射电子显微镜（TEM）证实，IGP蛋白存在于脂质体膜。在用IGP蛋白修饰后，将DOPC：DOPG脂质体应用于癌细胞。显微镜和流式细胞仪显示修饰的脂质体选择性地结合整合素αvβ3阳性的A549细胞。此外，与常用的化学偶联法相比，该方法在样品制备简单、成本低廉等方面具有上级优势。接下来，将IGP蛋白应用于修饰红细胞（RBC）脂质体，用于体外和体内应用中的靶向递送。IGP修饰的RBC脂质体优先靶向表达整合素αvβ3的A549癌细胞。体内成像显示，IGP修饰的RBC脂质体集中在肿瘤组织中，并主要由肝脏和肾脏代谢。
Antimicrobial peptides (AMPs) are typical cell penetrating peptides (CPPs) that intercalate into biomembranes and exhibit broad activities. We designed a triple fusion protein consisting of an AMP, Ib-AMP4 at the N-terminus, a fluorescent GFP probe in the center, and the tumor-targeting peptide P1c at the other terminus. After purification fromE. coli, the interaction between the Ib-AMP4-GFP-P1c fusion protein (IGP) and the lipid membrane was characterized. Experiments using isothermal titration calorimetry (ITC) and quartz crystal microbalance with dissipation (QCM-D) demonstrated that IGP proteins spontaneously bound the lipid bilayer with a maximal molar ratio of 1:52 (protein:lipid). Furthermore, transmission electron microscopy (TEM) confirmed that the IGP protein was present in the liposome membrane. After decoration with IGP proteins, the DOPC:DOPG liposomes were applied to cancer cells. Microscopy and flow cytometry reveal that the decorated liposomes selectively bound integrin αvβ3-positive A549 cells. In addition, compared with the common chemical conjugation method, the reported method seemed to be superior in certain aspects, such as simple sample preparation and cost-effectiveness. Next, the IGP protein was applied to decorate red blood cell (RBC) liposomes for targeted delivery in both in vitro and in vivo applications. The IGP-decorated RBC liposomes preferentially targeted integrin αvβ3 expressing A549 cancer cells. The in vivo imaging showed that IGP-decorated RBC liposomes were concentrated in tumor tissue and were primarily metabolized by the liver and kidney.