Anderson Bambi D, Nakamura Takafumi, Russell Stephen J, Peng Kah-Whye
Molecular Medicine Program, Mayo Clinic Cancer Center, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA.
Cancer Res. 2004 Jul 15;64(14):4919-26. doi: 10.1158/0008-5472.CAN-04-0884.
Live attenuated Edmonston B strain of measles virus (MV-Edm) is a potent and specific oncolytic agent, but the mechanism underlying its tumor selectivity is unknown. The virus causes cytopathic effects (CPEs) of extensive syncytial formation in tumor cells but minimal damage or cell killing in normal cells. The CPE is dependent on expression of viral proteins and the presence of CD46, the major cellular receptor of MV-Edm. Using a virally encoded soluble marker peptide to provide a quantitative readout of the level of viral gene expression, we determined that tumor cells and normal cells expressed comparable levels of viral proteins. CD46 mediates virus attachment, entry, and virus-induced cell-to-cell fusion. Using engineered cells expressing a range of CD46 densities, we determined that whereas virus entry increased progressively with CD46 density, cell fusion was minimal at low receptor densities but increased dramatically above a threshold density of CD46 receptors. It is well established that tumor cells express abundant CD46 receptors on their surfaces compared with their normal counterparts. Thus, at low CD46 densities typical of normal cells, infection occurs, but intercellular fusion is negligible. At higher densities typical of tumor cells, infection leads to extensive cell fusion. Intercellular fusion also results in enhancement of viral gene expression through recruitment of neighboring uninfected cells into the syncytium, further amplifying the CPE. Discrimination between high and low CD46 receptor density provides a compelling basis for the oncolytic specificity of MV-Edm and establishes MV-Edm as a promising CD46-targeted cancer therapeutic agent.
麻疹病毒(MV-Edm)的减毒活Edmonston B株是一种强效且特异的溶瘤剂,但其肿瘤选择性的潜在机制尚不清楚。该病毒在肿瘤细胞中会引起广泛的合胞体形成的细胞病变效应(CPE),但在正常细胞中造成的损伤或细胞杀伤极小。CPE依赖于病毒蛋白的表达以及MV-Edm的主要细胞受体CD46的存在。我们使用病毒编码的可溶性标记肽来定量读出病毒基因表达水平,确定肿瘤细胞和正常细胞表达的病毒蛋白水平相当。CD46介导病毒附着、进入以及病毒诱导的细胞间融合。通过使用表达一系列CD46密度的工程细胞,我们确定虽然病毒进入随着CD46密度的增加而逐渐增加,但在低受体密度下细胞融合极少,但在CD46受体密度超过阈值时急剧增加。与正常细胞相比,肿瘤细胞表面表达丰富的CD46受体,这一点已得到充分证实。因此,在正常细胞典型的低CD46密度下,会发生感染,但细胞间融合可忽略不计。在肿瘤细胞典型的较高密度下,感染会导致广泛的细胞融合。细胞间融合还通过将相邻未感染细胞募集到合胞体中导致病毒基因表达增强,进一步放大CPE。对高和低CD46受体密度的区分是MV-Edm溶瘤特异性的有力基础,并将MV-Edm确立为一种有前景的靶向CD46的癌症治疗剂。