Nanobody-Enhanced Targeting of AAV Gene Therapy Vectors
Nanobody-Enhanced Targeting of AAV Gene Therapy Vectors
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DOI：
10.1016/j.omtm.2019.09.003
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发表时间：
2019-12-13
影响因子：
4.7
通讯作者：
Koch-Nolte, Friedrich
中科院分区：
文献类型：
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作者：
Eichhoff, Anna Marei;Boerner, Kathleen;Koch-Nolte, Friedrich
在基因治疗中使用腺相关病毒（AAV）作为载体的限制因素是AAV血清型的广泛嗜性，即，几种细胞类型的平行感染。纳米抗体是来自骆驼科动物中天然存在的重链抗体的单个免疫球蛋白可变结构域。它们的小尺寸和高溶解度允许容易地重组成融合蛋白。在本文中，我们表明膜蛋白特异性纳米抗体可以插入到AAV 2的VP 1衣壳蛋白的表面环中。使用三种结构不同的膜蛋白-多跨离子通道，单跨膜蛋白，和糖基磷脂酰肌醇（GPI）锚定的胞外酶-我们表明，这种策略可以显着增强特定的靶细胞的转导重组腺相关病毒2。此外，我们显示AAV 2的纳米抗体-VP 1融合物可以掺入AAV 1、AAV 8和AAV 9的衣壳中，从而有效地重定向其他AAV血清型的靶特异性。纳米抗体介导的靶向提供了一种高效的AAV靶向策略，可能为细胞的基因工程开辟新的途径。
A limiting factor for the use of adeno-associated viruses (AAVs) as vectors in gene therapy is the broad tropism of AAV serotypes, i.e., the parallel infection of several cell types. Nanobodies are single immunoglobulin variable domains from heavy chain antibodies that naturally occur in camelids. Their small size and high solubility allow easy reformatting into fusion proteins. Herein we show that a membrane protein-specific nanobody can be inserted into a surface loop of the VP1 capsid protein of AAV2. Using three structurally distinct membrane proteins-a multispan ion channel, a single-span transmembrane protein, and a glycosylphosphatidylinositol (GPI)-anchored ectoenzyme-we show that this strategy can dramatically enhance the transduction of specific target cells by recombinant AAV2. Moreover, we show that the nanobody-VP1 fusion of AAV2 can be incorporated into the capsids of AAV1, AAV8, and AAV9 and thereby effectively redirect the target specificity of other AAV serotypes. Nano-body-mediated targeting provides a highly efficient AAV targeting strategy that is likely to open up new avenues for genetic engineering of cells.