Modulation of adenovirus vector tropism via incorporation of polypeptide ligands into the fiber protein
Modulation of adenovirus vector tropism via incorporation of polypeptide ligands into the fiber protein
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DOI：
10.1128/jvi.76.17.8621-8631.2002
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发表时间：
2002-09-01
影响因子：
5.4
通讯作者：
Krasnykh, V
中科院分区：
文献类型：
--
作者：
Belousova, N;Krendelchtchikova, V;Krasnykh, V
如果能够开发出能够组织特异性基因传递的病毒载体，基于腺病毒(Ad)的基因治疗的效果可能会显著提高。以前对Ad载体的取向进行基因修饰的尝试只取得了部分成功，这主要是由于可以整合到Ad衣壳中的配体的数量有限。早期的研究发现，Ad纤维蛋白的结构对其羧基末端的配体施加了严格的大小限制，从而将潜在的候选配体的范围限制在短肽上。我们先前已经确定纤维结节结构域的HI环是掺入靶向配体的首选位置，并假设该环的结构特性将允许插入各种配体，包括大的多肽分子。在目前的研究中，我们通过推导出一族其纤维含有长度递增的多肽插入物的Ad载体来检验这一假设。通过评估病毒的产量和感染性水平以及受体的特异性，我们发现，超过50%的结节结构域大小的多肽序列可以被整合到纤维中，而对载体的这些关键属性只会产生轻微的负面影响。我们的研究还揭示了用于载体修饰的配体的大小与所产生的病毒的感染性和产量之间的负相关，从而预测了进一步扩大纤维结节与病毒粒子的完整性不相容的限度。
The efficacy of adenovirus (Ad)-based gene therapy might be significantly improved if viral vectors capable of tissue-specific gene delivery could be developed. Previous attempts to genetically modify the tropism of Ad vectors have been only partially successful, largely due to the limited repertoire of ligands that can be incorporated into the Ad capsid. Early studies identified stringent size limitations imposed by the structure of the Ad fiber protein on ligands incorporated into its carboxy terminus and thus limited the range of potential ligand candidates to short peptides. We have previously identified the HI loop of the fiber knob domain as a preferred site for the incorporation of targeting ligands and hypothesized that the structural properties of this loop would allow for the insertion of a wide variety of ligands, including large polypeptide molecules. In the present study we have tested this hypothesis by deriving a family of Ad vectors whose fibers contain polypeptide inserts of incrementally increasing lengths. By assessing the levels of productivity and infectivity and the receptor specificities of the resultant viruses, we show that polypeptide sequences exceeding by 50% the size of the knob domain can be incorporated into the fiber with only marginal negative consequences on these key properties of the vectors. Our study has also revealed a negative correlation between the size of the ligand used for vector modification and the infectivity and yield of the resultant virus, thereby predicting the limits beyond which further enlargement of the fiber knob would not be compatible with the virion's integrity.