Novel targeted adenovirus

新型靶向腺病毒

基本信息

批准号：
9511780
负责人：
David Terry Curiel
金额：
$ 34.88万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-19 至 2022-05-31
项目状态：
已结题

项目摘要

ABSTRACT A central mandate to realize effective gene therapy is the ability to accomplish cell specific delivery. Capitalizing on the in vivo delivery efficacy of adenoviral vectors (Ad), recent studies have highlighted the capacity of targeted Ad to accomplish cell selectivity in this stringent delivery context. Systemic employment of Ad for gene delivery, however, is currently limited by vector particle sequestration in the liver. Recent work in several laboratories, however, has identified the biologic dictates of vector hepatotropism. Based on this understanding, it has now been possible to "un-target" the liver thereby facilitating strategies designed to achieve cell specific gene delivery in the context of systemic vector administration. Of note, we have recently shown that such liver un-targeting strategies can synergize with described vector targeting methods such as those based upon restricting delivered transgene expression to target cells with a tissue/tumor selective promoter ("transcriptional targeting"). The dramatic synergistic specificity gains noted with combination of these two approaches logically suggests that further gains may accrue additional targeting methods exploited in combination. In this regard, strategies have been proposed to target Ad based upon re-directing vector binding to target cell specific cell surface markers. Such "transductional" targeting methods would offer potential synergies with the targeting methods we note above. Such an endeavor has been limited to this point by the inability of current vector engineering to achieve capsid incorporation of antibody targeting species. Herein we seek to address this key limit. First, we have developed a method to replace the native adenovirus fiber with a substitute chimera devoid of the native fiber's knob binding domain. This maneuver eliminates native tropism and allows for the incorporation of a wider range of large/complex candidate targeting ligands. Second, we have demonstrated that the single domain antibody species derived from camelids (sdAb) possess the unique attributes allowing biologic compatibility with adenovirus capsid synthesis and assembly. In the aggregate, these two technologies now allow for the functional incorporation of antibody targeting species into the Ad capsid for the achievement of cell-specific targeting. This additional level of targeting provides for potential synergies with the defined liver un-targeting and transcriptional vector targeting methods we have explored.

抽象的实现有效基因疗法的核心任务是完成特定细胞的能力送货。利用腺病毒载体（AD）的体内递送功效，最近的研究强调了目标广告在此严格的情况下实现细胞选择性的能力交付环境。但是，目前，系统地就基因递送的广告就业受到了限制肝脏中的矢量颗粒隔离。然而，最近在几个实验室的工作已经确定了载体肝动体的生物学决定。基于这种理解，它现在有有可能“取消目标”肝脏，从而促进旨在实现细胞的策略在系统媒介给药的背景下的特定基因传递。值得注意的是，我们最近有表明这种肝脏不靶向策略可以与描述的向量靶向协同作用诸如基于限制传递的转基因表达到目标细胞的方法与组织/肿瘤选择性启动子（“转录靶向”）。戏剧性的协同作用通过这两种方法的结合在逻辑上提出的特异性获得了，这进一步表明增益可能会产生组合利用的其他目标方法。在这方面，已经提出了基于重新导向矢量结合到目标细胞的策略来靶向AD 特定的细胞表面标记。这种“转移”靶向方法将提供潜力我们在上面注意到的定位方法协同作用。这样的努力仅限于此通过当前矢量工程无法实现抗体的capsID掺入的点靶向物种。在此，我们试图解决此关键限制。首先，我们开发了一种方法用替代无原生纤维的替代嵌合体代替本地腺病毒纤维结合域。这种操纵消除了本地的偏向主义，并允许合并大/复杂候选配体的范围更宽。其次，我们已经证明了来自骆驼（SDAB）的单域抗体物种具有独特的属性允许生物学兼容性与腺病毒衣壳合成和组装。在总体中，这两种技术现在允许靶向抗体的功能掺入进入AD CAPSID，以实现细胞特异性靶向。这个额外的定位水平提供了与定义的肝脏和转录向量的潜在协同作用我们探索的定位方法。