Targeting Adenoviruses with Factor X-Single-Chain Antibody Fusion Proteins
Targeting Adenoviruses with Factor X-Single-Chain Antibody Fusion Proteins
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DOI：
10.1089/hum.2009.190
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发表时间：
2010-06-01
影响因子：
4.2
通讯作者：
Barry, Michael A.
中科院分区：
文献类型：
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作者：
Chen, Christopher Y.;May, Shannon M.;Barry, Michael A.
研究表明，包括因子 X (FX) 在内的凝血因子可与腺病毒血清型 5 (Ad5) 六邻体蛋白结合，并在静脉注射后将病毒靶向肝细胞。这些因子通过其保守的维生素 K 依赖性 γ-羧基谷氨酸 (GLA) 结构域以亚纳摩尔亲和力与六邻体结合。在这项工作中，我们利用与单链抗体可变片段 (ScFv) 融合的 FX 的 GLA 结构域，利用这种强相互作用将 Ad 重新定位到新受体。我们证明，人 FX 的 GLA 结构域与受体特异性 ScFv 的融合将使 Ad5 载体靶向表达这些受体的细胞。与非靶向病毒相比，α Her2 ScFv 与 GLA 的融合增加了 Her2 阳性细胞与 Her2 阴性细胞的体外转导。针对表皮生长因子受体 (EGFR) 和干细胞标记 ATP 结合盒蛋白 G2 (ABCG2) 的 ScFv 也获得了类似的结果。复制缺陷型或复制型 Ad 的 GLA 融合蛋白的直接表达增加了体外和体内癌细胞的感染和杀伤。这些数据证明了使用 GLA 结构域桥接分泌配体与细胞内产生的 Ad5 载体以实现载体靶向的潜力。
It has been shown that blood clotting factors, including factor X (FX), bind to the adenovirus serotype 5 (Ad5) hexon protein and target the virus to liver hepatocytes after intravenous injection. These factors bind to hexon via their conserved vitamin K-dependent gamma-carboxyglutamic acid (GLA) domains with subnanomolar affinity. In this work, we have used this strong interaction to retarget Ad to new receptors, using the GLA domain of FX fused to single-chain antibody variable fragment (ScFv). We demonstrate that fusion of the GLA domain of human FX to receptor-specific ScFvs will target Ad5 vectors to cells expressing these receptors. Fusion of an alpha Her2 ScFv to GLA increased in vitro transduction of Her2-positive versus Her2-negative cells when compared with untargeted virus. Similar results were obtained with ScFvs against the epidermal growth factor receptor (EGFR) and against the stem cell marker ATP-binding cassette protein G2 (ABCG2). Direct expression of GLA fusion protein from replication-defective or replication-competent Ad increased infection and killing of cancer cells in vitro and in vivo. These data demonstrate the potential of using GLA domains to bridge secreted ligands with intracellularly produced Ad5 vectors for vector targeting.