Adenoviral vectors coated with PAMAM dendrimer conjugates allow CAR independent virus uptake and targeting to the EGF receptor.
Adenoviral vectors coated with PAMAM dendrimer conjugates allow CAR independent virus uptake and targeting to the EGF receptor.
复制标题
DOI：
10.1021/mp300366f
复制
发表时间：
2013-01
影响因子：
4.9
通讯作者：
Alexandra Vetter;K. Virdi;Sigrid Espenlaub;W. Rödl;E. Wagner;P. Holm;C. Scheu;F. Kreppel;C. Spitzweg;M. Ogris
中科院分区：
文献类型：
--
作者：
Alexandra Vetter;K. Virdi;Sigrid Espenlaub;W. Rödl;E. Wagner;P. Holm;C. Scheu;F. Kreppel;C. Spitzweg;M. Ogris
5型腺病毒（Adenovirus type 5，Ad）是一种高效的基因载体，具有很高的基因转导潜力，但其效率依赖于其天然细胞受体科萨基和腺病毒受体（CAR）的细胞粘附和α（v）β（3/5）整合素的内化。为了能够转导CAR阴性癌细胞系，我们已经通过与阳离子PAMAM（聚酰胺胺）树状聚合物的非共价电荷相互作用来包被带负电荷的Ad。通过使用肽配体GE 11将包被的Ad靶向表皮生长因子受体，增加了肿瘤细胞感染的特异性，GE 11通过2 kDa的PEG间隔基与PAMAM树枝状聚合物偶联。通过测量表面电荷和尺寸来检查颗粒，通过透射电子显微镜来确定包覆程度。PAMAM包被的Ad的净正电荷增强了细胞结合和摄取，导致转导效率增加，特别是在使用增强的绿色荧光蛋白或荧光素酶作为转基因的低至中等CAR表达癌细胞系中。虽然PAMAM包被的Ad允许有效的内化，但用线性聚乙烯亚胺包被诱导了过度的颗粒聚集，提高了细胞毒性并降低了转导效率。PAMAM包被的Ad能够成功的细胞在体外转导，即使在中和抗体的存在下。总之，这项研究清楚地证明了非共价的，基于电荷的包被的Ad载体与配体装备的树枝状聚合物作为一种可行的策略，有效转导细胞，否则难治性的Ad感染。
Adenovirus type 5 (Ad) is an efficient gene vector with high gene transduction potential, but its efficiency depends on its native cell receptors coxsackie- and adenovirus receptor (CAR) for cell attachment and α(v)β(3/5) integrins for internalization. To enable transduction of CAR negative cancer cell lines, we have coated the negatively charged Ad by noncovalent charge interaction with cationic PAMAM (polyamidoamine) dendrimers. The specificity for tumor cell infection was increased by targeting the coated Ad to the epidermal growth factor receptor using the peptide ligand GE11, which was coupled to the PAMAM dendrimer via a 2 kDa PEG spacer. Particles were examined by measuring surface charge and size, the degree of coating was determined by transmission electron microscopy. The net positive charge of PAMAM coated Ad enhanced cellular binding and uptake leading to increased transduction efficiency, especially in low to medium CAR expressing cancer cell lines using enhanced green fluorescent protein or luciferase as transgene. While PAMAM coated Ad allowed for efficient internalization, coating with linear polyethylenimine induced excessive particle aggregation, elevated cellular toxicity and lowered transduction efficiency. PAMAM coating of Ad enabled successful transduction of cells in vitro even in the presence of neutralizing antibodies. Taken together, this study clearly proves noncovalent, charge-based coating of Ad vectors with ligand-equipped dendrimers as a viable strategy for efficient transduction of cells otherwise refractory to Ad infection.