Grill Jacques, Lamfers Martine L M, van Beusechem Victor W, Dirven Clemens M, Pherai D Shareen, Kater Mathijs, Van der Valk Paul, Vogels Ronald, Vandertop W Peter, Pinedo Herbert M, Curiel David T, Gerritsen Winald R
Division of Gene Therapy, Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Mol Ther. 2002 Nov;6(5):609-14.
The use of adenoviruses for gene transfer and as oncolytic agents is currently receiving widespread attention. As specific constraints to adenovirus distribution and spread cannot be studied in cell cultures, there is a need for an in vitro three-dimensional (3D) model mimicking the in vivo biology of tumors. We studied the interactions between tumor and adenoviruses using multicellular spheroids grown from primary brain tumor material. Using beta-galactosidase and luciferase reporter genes expressed by replication-defective adenoviruses, we showed that infection was restricted to the first layer of cells. Using a replication-competent adenovirus expressing the luciferase gene, we showed that transgene expression in the spheroid was considerably enhanced and that viral spreading deep into the 3D structure took place. In addition, a tetrazolium salt-based metabolic assay could be used to compare the oncolytic activity of different concentrations of replication-competent adenoviruses. We can conclude that organotypic spheroids offer a versatile in vitro system for studying distribution, spread, and oncolysis by adenoviruses in a clinically relevant model.
目前，腺病毒在基因传递及作为溶瘤药物方面的应用正受到广泛关注。由于无法在细胞培养中研究腺病毒分布和传播的特定限制因素，因此需要一种体外三维（3D）模型来模拟肿瘤的体内生物学特性。我们使用从原发性脑肿瘤材料生长的多细胞球体研究肿瘤与腺病毒之间的相互作用。利用复制缺陷型腺病毒表达的β-半乳糖苷酶和荧光素酶报告基因，我们发现感染仅限于细胞的第一层。使用表达荧光素酶基因的复制型腺病毒，我们发现球体中的转基因表达显著增强，并且病毒深入三维结构传播。此外，基于四唑盐的代谢测定可用于比较不同浓度复制型腺病毒的溶瘤活性。我们可以得出结论，器官型球体为在临床相关模型中研究腺病毒的分布、传播和溶瘤作用提供了一个通用的体外系统。
