The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, Department of Biomedicine & Health Sciences, The Catholic University of Korea, Seoul, 06591, Republic of Korea.
am SCIENCES, C-912, SK V1 GL Metrocity, 128, Beobwonro, Songpa-gu, Seoul, 05854, Republic of Korea.
Sci Rep. 2019 Feb 7;9(1):1616. doi: 10.1038/s41598-019-38533-7.
Virus-like particles (VLPs) possess great potential for organ-specific transport of therapeutic agents due to their central cavity surrounded by viral capsid proteins and similar tropism to their original viruses. The N-terminal truncated second open reading frame (Nt-ORF2) of the hepatotropic hepatitis E virus (HEV) forms VLPs via self-assembly. In the present study, we investigated whether HEV-LPs could deliver foreign genes specifically to the liver. HEV-LPs were obtained from Nt-ORF2 expression in Huh7 cells that were transduced with recombinant baculoviruses and purified by continuous density gradient centrifugation. The purified HEV-LPs efficiently penetrated liver-derived cell lines and the liver tissues. To evaluate HEV-LPs as gene delivery tools, we encapsulated foreign plasmids in HEV-LPs with disassembly/reassembly systems. Green fluorescence was detected at higher frequency in liver-derived Huh7 cells treated with HEV-LPs bearing GFP-encoding plasmids than in control cells. Additionally, HEV-LPs bearing Bax-encoding plasmids induced apoptotic signatures in Huh7 cells. In conclusion, HEV-LPs produced in mammalian cells can encapsulate foreign genes in their central cavity and specifically transport these genes to liver-derived cells, where they are expressed. The present study could contribute to advances in liver-targeted gene therapy.
病毒样颗粒（VLPs）由于其中央腔被病毒衣壳蛋白包围，并且具有与原始病毒相似的趋向性，因此具有将治疗剂靶向递送至特定器官的巨大潜力。嗜肝性肝炎 E 病毒（HEV）的 N 端截断的第二个开放阅读框（Nt-ORF2）通过自组装形成 VLPs。在本研究中，我们研究了 HEV-LPs 是否可以将外源基因特异性递送至肝脏。通过转导重组杆状病毒并通过连续密度梯度离心纯化，从 Huh7 细胞中转录的 Nt-ORF2 表达中获得了 HEV-LPs。纯化的 HEV-LPs 能够有效地穿透肝源性细胞系和肝组织。为了评估 HEV-LPs 作为基因传递工具的作用，我们使用拆卸/重新组装系统将外源质粒封装在 HEV-LPs 中。与用携带 GFP 编码质粒的 HEV-LPs 处理的对照细胞相比，用携带 GFP 编码质粒的 HEV-LPs 处理的肝源性 Huh7 细胞中检测到绿色荧光的频率更高。此外，携带 Bax 编码质粒的 HEV-LPs 在 Huh7 细胞中诱导了凋亡特征。总之，在哺乳动物细胞中产生的 HEV-LPs 可以将外源基因封装在其中央腔中，并将这些基因特异性递送至肝源性细胞，在这些细胞中表达。本研究可能有助于推进肝靶向基因治疗。
