Adenoviral gene vector tethering to nanoparticle surfaces results in receptor-independent cell entry and increased transgene expression
Adenoviral gene vector tethering to nanoparticle surfaces results in receptor-independent cell entry and increased transgene expression
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DOI：
10.1016/j.ymthe.2006.03.023
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发表时间：
2006-09-01
影响因子：
12.4
通讯作者：
Levy, Robert J.
中科院分区：
文献类型：
--
作者：
Chorny, Michael;Fishbein, Ilia;Levy, Robert J.
本研究探讨了一种假设，即在可生物降解纳米颗粒（NP）表面亲和固定复制缺陷腺病毒（Ad）可以通过解耦柯萨奇-腺病毒受体（CAR）的细胞摄取来改善转导。用光反应性聚烯丙胺-二苯甲酮-吡啶二硫代羧酸聚合物对聚乳酸基NP进行表面活化，使Ad结合蛋白（重组CAR的D1结构域或腺病毒球形特异性单克隆抗体）能够（通过硫醇化学）共价附着。研究了NP-Ad复合物的基因转移与细胞摄取、细胞类型和NP-Ad结合特性的关系。NP-Ad复合物，而非Ad与对照非免疫igg修饰的NP一起或不一起应用，在内皮瘤和内皮和动脉平滑肌细胞（SMC）中显著增加绿色荧光蛋白报告蛋白的表达，这与NP-Ad内化的程度直接相关。通过显示抑制游离Ad而不是旋钮蛋白介导的NP-Ad复合物的转导，证实了NP-Ad的car非依赖性摄取。与含有（GFP）Ad或（NULL）Ad或自由载体的配合物相比，含有编码诱导型一氧化氮合酶的Ad配制的配合物对培养SMC生长的抑制作用明显更大。综上所述，ad特异性亲和性粘附于可生物降解的NIP可以通过不依赖car的摄取机制显著提高基因表达水平。
The present studies investigated the hypothesis that affinit4y immobilization of replication-defective adenoviruses (Ad) on the surfaces of biodegradable nanoparticles (NP) can improve transduction through uncoupling cellular uptake from the coxsackie-adenovirus receptor (CAR). Ad was tethered to the surfaces of polylactide-based NP that were surface-activated using a photoreactive polyallylamine-benzophenone-pyridyidithiocarboxylate polymer, which enabled (via thiol chemistry) the covalent attachment of Ad-binding proteins, either the recombinant D1 domain of CAR or an adenoviral knob-specific monoclonal antibody. Gene transfer by NP-Ad complexes was studied in relation to cellular uptake as a function of cell type and the character of NP-Ad binding. NP-Ad complexes, but not Ad applied with or without control nonimmune IgG-modified NP, significantly increased green fluorescent protein reporter expression in endothelioma and endothelial and arterial smooth muscle cells (SMC) in direct correlation to the extent of NP-Ad internalization. CAR-independent uptake of NP-Ad was confirmed by demonstrating inhibition of free Ad- but not NP-Ad complex-mediated transduction by knob protein. Complexes formulated with an Ad encoding inducible nitric oxide synthase inhibited growth of cultured SMC to a significantly greater extent than those with (GFP)Ad or (NULL)Ad or free vector. It is concluded that Ad-specific affinity tethering to biodegradable NIP can significantly increase the level of gene expression via a CAR-independent uptake mechanism.