The AAV9 receptor and its modification to improve in vivo lung gene transfer in mice
The AAV9 receptor and its modification to improve in vivo lung gene transfer in mice
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DOI：
10.1172/jci57367
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发表时间：
2011-06-01
影响因子：
15.9
通讯作者：
Wilson, James M.
中科院分区：
文献类型：
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作者：
Bell, Christie L.;Vandenberghe, Luk H.;Wilson, James M.
基于腺相关病毒（AAV）血清型9的载体是体内基因递送至许多器官的候选者，但介导这些向性的受体尚未被定义。我们评估了具有末端唾液酸（SA）的聚糖对AAV 9的摄取，这是病毒进入细胞的常见模式。然而，我们发现，当末端SA被酶促去除时，AAV 9结合增加，这表明半乳糖（其是SA的最常见的倒数第二单糖）可能介导AAV 9转导。这在糖蛋白生物合成相关酶缺陷的突变CHO Pro-5细胞以及凝集素干扰研究中得到证实。通过聚糖结合测定证明了AAV 9与具有末端半乳糖的聚糖的结合。将AAV 9载体与神经氨酸酶共滴注到小鼠肺中导致末端半乳糖暴露在传导气道上皮细胞的顶端表面上，如通过凝集素结合和这些细胞的转导增加所示，证明了该载体在肺定向基因转移中的可能效用。增加靶细胞上受体的丰度和改善载体功效可以改善AAV载体向其治疗靶标的递送。
Vectors based on adeno-associated virus (AAV) serotype 9 are candidates for in vivo gene delivery to many organs, but the receptor(s) mediating these tropisms have yet to be defined. We evaluated AAV9 uptake by glycans with terminal sialic acids (SAs), a common mode of cellular entry for viruses. We found, however, that AAV9 binding increased when terminal SA was enzymatically removed, suggesting that galactose, which is the most commonly observed penultimate monosaccharide to SA, may mediate AAV9 transduction. This was confirmed in mutant CHO Pro-5 cells deficient in the enzymes involved in glycoprotein biogenesis, as well as lectin interference studies. Binding of AAV9 to glycans with terminal galactose was demonstrated via glycan binding assays. Co-instillation of AAV9 vector with neuraminidase into mouse lung resulted in exposure of terminal galactose on the apical surface of conducting airway epithelial cells, as shown by lectin binding and increased transduction of these cells, demonstrating the possible utility of this vector in lung-directed gene transfer. Increasing the abundance of the receptor on target cells and improving vector efficacy may improve delivery of AAV vectors to their therapeutic targets.