Construction of a Fully Retargeted Herpes Simplex Virus 1 Recombinant Capable of Entering Cells Solely via Human Epidermal Growth Factor Receptor 2
Construction of a Fully Retargeted Herpes Simplex Virus 1 Recombinant Capable of Entering Cells Solely via Human Epidermal Growth Factor Receptor 2
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DOI：
10.1128/jvi.01133-08
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发表时间：
2008-10-01
影响因子：
5.4
通讯作者：
Campadelli-Fiume, Gabriella
中科院分区：
文献类型：
--
作者：
Menotti, Laura;Cerretani, Arianna;Campadelli-Fiume, Gabriella
一个新的前沿在治疗肿瘤是难以治疗的溶瘤病毒疗法，其中复制能力的病毒选择性地感染和破坏肿瘤细胞。单纯疱疹病毒（HSV）是一个特别有吸引力的系统。已经通过在gD中插入异源配体实现了对肿瘤特异性受体的有效重靶向。以前，我们的实验室产生了一种重靶向人表皮生长因子受体2（HER 2）的HSV，该受体在约三分之一的乳腺肿瘤和一些卵巢肿瘤中过表达。HER 2过表达与转移性增加和预后不良相关。由于HER 2没有天然配体，因此插入的配体是HER 2的单链抗体。这项工作的目的是对HSV进行遗传工程改造，使其选择性靶向表达HER 2的肿瘤细胞，并且已经失去了通过天然gD受体HVEM和nectin 1进入细胞的能力。通过两种不同的策略，点突变和在不同于先前描述的插入位点的gD中的位点处插入单链抗体，尝试从nectin 1去靶向。我们报告了gD氨基酸34、215、222和223处的点突变未能产生nectin 1去靶向HSV。通过将单链抗体插入位点移动到残基39，即，在nectin 1相互作用表面的前面，而不是在它的侧面，并通过删除氨基酸残基6至38。所得到的重组体R-LM 113进入细胞并仅通过HER 2在细胞间扩散。
A novel frontier in the treatment of tumors that are difficult to treat is oncolytic virotherapy, in which a replication-competent virus selectively infects and destroys tumor cells. Herpes simplex virus (HSV) represents a particularly attractive system. Effective retargeting to tumor-specific receptors has been achieved by insertion in gD of heterologous ligands. Previously, our laboratory generated an HSV retargeted to human epidermal growth factor receptor 2 (HER2), a receptor overexpressed in about one-third of mammary tumors and in some ovarian tumors. HER2 overexpression correlates with increased metastaticity and poor prognosis. Because HER2 has no natural ligand, the inserted ligand was a single-chain antibody to HER2. The objective of this work was to genetically engineer an HSV that selectively targets the HER2-expressing tumor cells and that has lost the ability to enter cells through the natural gD receptors, HVEM and nectin1. Detargeting from nectin1 was attempted by two different strategies, point mutations and insertion of the single-chain antibody at a site in gD different from previously described sites of insertion. We report that point mutations at gD amino acids 34, 215, 222, and 223 failed to generate a nectin1-detargeted HSV. An HSV simultaneously detargeted from nectin1 and HVEM and retargeted to HER2 was successfully engineered by moving the site of single-chain antibody insertion at residue 39, i.e., in front of the nectin1-interacting surface and not lateral to it, and by deleting amino acid residues 6 to 38. The resulting recombinant, R-LM113, entered cells and spread from cell to cell solely via HER2.