Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors
Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors
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DOI：
10.1038/ncomms7246
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发表时间：
2015-02-01
影响因子：
16.6
通讯作者：
Buchholz, Christian J.
中科院分区：
文献类型：
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作者：
Muench, Robert C.;Muth, Anke;Buchholz, Christian J.
我们描述了受体靶向的腺相关病毒（AAV）载体，其允许离体和体内罕见细胞类型的遗传修饰，同时显示出不可检测的脱靶。通过在病毒衣壳上重新设计锚蛋白重复蛋白（DARPin）并小心地耗尽DARPin缺陷颗粒，制备对Her 2/neu、EpCAM或CD 4特异的AAV载体。靶向肿瘤抗原Her 2/neu的载体的单次静脉内施用足以追踪75%的所有肿瘤部位，并且比细胞抑制抗体Herceptin延长存活时间。靶向CD 4的AAV击中人源化小鼠模型的脾脏中存在的人CD 4阳性细胞，而CD 8阳性细胞以及肝脏或其他脱靶器官保持未修饰。EpCAM-AAV模拟循环肿瘤细胞的条件，检测到人类血液中的单个肿瘤细胞，为肿瘤干细胞追踪开辟了道路。因此，这里开发的方法将基因递送到具有抗体样特异性的选择的靶细胞类型。
We describe receptor-targeted adeno-associated viral (AAV) vectors that allow genetic modification of rare cell types ex vivo and in vivo while showing no detectable off-targeting. Displaying designed ankyrin repeat proteins (DARPins) on the viral capsid and carefully depleting DARPin-deficient particles, AAV vectors were made specific for Her2/neu, EpCAM or CD4. A single intravenous administration of vector targeted to the tumour antigen Her2/neu was sufficient to track 75% of all tumour sites and to extend survival longer than the cytostatic antibody Herceptin. CD4-targeted AAVs hit human CD4-positive cells present in spleen of a humanized mouse model, while CD8-positive cells as well as liver or other off-target organs remained unmodified. Mimicking conditions of circulating tumour cells, EpCAM-AAV detected single tumour cells in human blood opening the avenue for tumour stem cell tracking. Thus, the approach developed here delivers genes to target cell types of choice with antibody-like specificity.