A hepatitis B virus-derived human hepatic cell-specific heparin-binding peptide: identification and application to a drug delivery system
A hepatitis B virus-derived human hepatic cell-specific heparin-binding peptide: identification and application to a drug delivery system
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DOI：
10.1039/c8bm01134f
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发表时间：
2019-01-01
影响因子：
6.6
通讯作者：
Kuroda, Shun'ichi
中科院分区：
文献类型：
--
作者：
Liu, Qiushi;Somiya, Masaharu;Kuroda, Shun'ichi
病毒是自然进化的纳米载体，可以逃避宿主免疫系统，特异性地附着在靶细胞表面，通过内吞作用进入细胞，有效地从内体逃逸，然后将其基因组转移到宿主细胞。B型肝炎病毒（Hepatitis B virus，HBV）是一种直径约42 nm的有包膜DNA病毒，可特异性感染人肝细胞。为了在合成纳米载体中利用HBV衍生的早期感染机制，人肝细胞结合位点（即，牛磺胆酸钠共转运多肽（NTCP）结合位点，与肉豆蔻酰化的前S1（2-47））和低pH依赖性融合结构域（前S1（9-24））分别是靶向和内体逃逸所必需的。然而，细胞表面NTCP最近已被证明不参与HBV的初始附着。在这项研究中，我们确定了一个新的肝素结合位点（pre-S1（30-42））在N-末端的前S1区的一半，推测与细胞表面硫酸乙酰肝素蛋白聚糖（HSPG）相互作用，并在HBV的初始附着到人肝细胞中起着关键作用。进化上保守的氨基酸残基Asp-31、Trp-32和Asp-33对于肝素结合活性是必不可少的。展示肽的脂质体（LP）以细胞表面肝素依赖性方式被人肝细胞内吞，并比肉豆蔻酰化的pre-S1（2-47）展示LP更有效地将阿霉素递送至人肝细胞。这些结果表明，pre-S1（30-42）肽是合成纳米载体的最有前途的HBV衍生靶向肽，并且该肽对人肝细胞表现出高特异性并有效地诱导内吞作用。
Viruses are naturally evolved nanocarriers that can evade host immune systems, attach specifically to the surfaces of target cells, enter the cells through endocytosis, escape from endosomes efficiently, and then transfer their genomes to host cells. Hepatitis B virus (HBV) is a approximate to 42 nm enveloped DNA virus that can specifically infect human hepatic cells. To utilize the HBV-derived early infection machinery in synthetic nanocarriers, the human hepatic cell-binding site (i.e., the sodium taurocholate co-transporting polypeptide (NTCP)-binding site, with myristoylated pre-S1(2-47)) and the low pH-dependent fusogenic domain (pre-S1(9-24)) are indispensable for targeting and endosomal escape, respectively. However, cell-surface NTCP has recently been shown not to be involved in the initial attachment of HBV. In this study, we identified a novel heparin-binding site (pre-S1(30-42)) in the N-terminal half of the pre-S1 region, which presumably interacts with cell-surface heparan sulfate proteoglycan (HSPG) and plays a pivotal role in the initial attachment of HBV to human hepatic cells. The evolutionarily conserved amino acid residues Asp-31, Trp-32, and Asp-33 are indispensable for the heparin-binding activity. Liposomes (LPs) displaying the peptide were endocytosed by human hepatic cells in a cell-surface heparin-dependent manner and delivered doxorubicin to human hepatic cells more efficiently than myristoylated pre-S1(2-47)-displaying LPs. These results demonstrated that the pre-S1(30-42) peptide is the most promising HBV-derived targeting peptide for synthetic nanocarriers, and that this peptide exhibits high specificity for human hepatic cells and efficiently induces endocytosis.