Reduction of natural adenovirus tropism to mouse liver by fiber-shaft exchange in combination with both CAR- and αv integrin-binding ablation
Reduction of natural adenovirus tropism to mouse liver by fiber-shaft exchange in combination with both CAR- and αv integrin-binding ablation
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DOI：
10.1128/jvi.77.24.13062-13072.2003
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发表时间：
2003-12-01
影响因子：
5.4
通讯作者：
Hayakawa, T
中科院分区：
文献类型：
--
作者：
Koizumi, N;Mizuguchi, H;Hayakawa, T
主要受体柯萨奇病毒和腺病毒受体(CAR)和次级受体αV整合素是5型腺病毒(Ad)的趋向性决定因素。抑制纤维与CAR和五肽碱基与Alphav整合素的相互作用对于靶向转导特定细胞群的Ad载体的发展至关重要。在这项研究中，我们通过突变五元碱基的纤维结节和RGD基序，开发了同时去除CAR和αV整合素结合的Ad载体。我们还用来自Ad类型35的结构域替换了纤维轴结构域。静脉注射双突变体Ad载体后，小鼠肝脏中的高转导效率被抑制了约130-270倍，与传统的Ad载体(5型)相比，该载体分别突变了五酮碱基的两个纤维节和轴以及RGD基序。最值得注意的是，含有去除CAR结合能力的纤维结节、Ad类型35的纤维轴和缺失RGD基序的五元碱基的三突变体Ad载体比传统的Ad载体介导的小鼠肝脏转导水平低30,000倍。这个三重突变的Ad载体还介导了其他器官(脾、肾、心脏和肺)转导的减少。病毒DNA分析表明，系统递送的三突变体Ad载体主要被肝脏非实质细胞摄取，大多数病毒DNA容易被降解，导致肝脏中几乎没有基因表达。这些结果表明，五肽碱基的纤维节、纤维轴和RGD基序在Ad载体介导的小鼠肝脏转导中起着重要作用，并且三个突变体的Ad载体对任何器官几乎没有趋向性，似乎是靶向Ad载体的基本载体。
The primary receptor, the coxsackievirus and adenovirus receptor (CAR), and the secondary receptor, alphav integrins, are the tropism determinants of adenovirus (Ad) type 5. Inhibition of the interaction of both the fiber with CAR and the penton base with the alphav integrin appears to be crucial to the development of targeted Ad vectors, which specifically transduce a given cell population. In this study, we developed Ad vectors with ablation of both CAR and alphav integrin binding by mutating the fiber knob and the RGD motif of the penton base. We also replaced the fiber shaft domain with that derived from Ad type 35. High transduction efficiency in the mouse liver was suppressed approximately 130- to 270-fold by intravenous administration of the double-mutant Ad vectors, which mutated two domains each of the fiber knob and shaft and the RGD motif of the penton base compared with those of conventional Ad vectors (type 5). Most significantly, the triple-mutant Ad vector containing the fiber knob with ablation of CAR binding ability, the fiber shaft of Ad type 35, and the penton base with a deletion of the RGD motif mediated a >30,000-fold lower level of mouse liver transduction than the conventional Ad vectors. This triple-mutant Ad vector also mediated reduced transduction in other organs (the spleen, kidney, heart, and lung). Viral DNA analysis showed that systemically delivered triple-mutant Ad vector was primarily taken up by liver nonparenchymal cells and that most viral DNAs were easily degraded, resulting in little gene expression in the liver. These results suggest that the fiber knob, fiber shaft, and RGD motif of the penton base each plays an important role in Ad vector-mediated transduction to the mouse liver and that the triple-mutant Ad vector exhibits little tropism to any organs and appears to be a fundamental vector for targeted Ad vectors.