The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting.
The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting.
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DOI：
10.1038/labinvest.2014.78
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发表时间：
2014-08
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血管内皮细胞（EC）是理想的基因治疗靶点，因为它们提供广泛的组织通路，并且是静脉内载体施用后的第一接触表面。人重组腺病毒血清型5（Ad 5）是最常用的基因转移系统，因为其可观的转基因负载能力和缺乏体细胞突变的风险。然而，标准的Ad 5载体在静脉内施用后主要覆盖肝脏而不是脉管系统。我们最近开发了一种将髓样细胞结合肽（MBP）整合到knob缺失的T4纤维蛋白嵌合纤维中的Ad 5载体（Ad.MBP），该载体可能通过载体传递机制来抑制肺EC。在这里，我们测试了Ad.MBP载体的全身向性、其骨髓细胞必要性和载体-EC表达剂量反应。利用多器官共免疫荧光分析，我们发现Ad.MBP在肺、心脏、肾脏、骨骼肌、胰腺、小肠和脑中产生广泛的EC转导。令人惊讶的是，Ad.MBP保留了肝细胞向性，尽管与标准Ad 5相比频率降低。虽然特异性结合骨髓细胞离体，多器官Ad.MBP表达不依赖于循环单核细胞或巨噬细胞。Ad.MBP剂量递减维持了完全的肺靶向能力，但显著降低了其他器官中的转基因表达。用CMV启动子/增强子替换EC特异性ROBO 4废除了肝细胞表达，但也降低了其他器官中的基因表达。总的来说，我们的多层次靶向策略可以在以前无法进入的器官中实现治疗性生物生产，这些器官会引发最具破坏性或致命性的人类疾病。
Vascular endothelial cells (ECs) are ideal gene therapy targets as they provide widespread tissue access and are the first contact surfaces following intravenous vector administration. Human recombinant adenovirus serotype 5 (Ad5) is the most frequently used gene transfer system because of its appreciable transgene payload capacity and lack of somatic mutation risk. However, standard Ad5 vectors predominantly transduce liver but not the vasculature following intravenous administration. We recently developed an Ad5 vector with a myeloid cell-binding peptide (MBP) incorporated into the knob-deleted, T4 fibritin chimeric fiber (Ad.MBP).This vector was shown to transduce pulmonary ECs presumably via a vector handoff mechanism. Here we tested the body wide tropism of the Ad.MBP vector, its myeloid cell necessity, and vector-EC expression dose response. Using comprehensive multi-organ co-immunofluorescence analysis, we discovered that Ad.MBP produced widespread EC transduction in lung, heart, kidney, skeletal muscle, pancreas, small bowel, and brain. Surprisingly, Ad.MBP retained hepatocyte tropism albeit at a reduced frequency compared with standard Ad5. While binding specifically to myeloid cells ex vivo, multi-organ Ad.MBP expression was not dependent on circulating monocytes or macrophages. Ad.MBP dose de-escalation maintained full lung targeting capacity but drastically reduced transgene expression in other organs. Swapping the EC-specific ROBO4 for the CMV promoter/enhancer abrogated hepatocyte expression but also reduced gene expression in other organs. Collectively, our multilevel targeting strategy could enable therapeutic biologic production in previously inaccessible organs that initiate the most debilitating or lethal human diseases.
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DOI：
10.1038/nrg3296
发表时间：
2012-11
期刊：
Nature reviews. Genetics
影响因子：
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作者：
通讯作者：
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影响因子：
5.4
作者：
Belousova, N;Korokhov, N;Krasnykh, V
通讯作者：
Krasnykh, V
影响因子：
11.2
作者：
Franses JW;Edelman ER
通讯作者：
Edelman ER
影响因子：
82.9
作者：
Cole, C;Qiao, J;Vile, RG
通讯作者：
Vile, RG
影响因子：
5.1
作者：
Alberti, M. O.;Deshane, J. S.;Roth, J. C.
通讯作者：
Roth, J. C.