Nakamura Takafumi, Peng Kah-Whye, Vongpunsawad Sompong, Harvey Mary, Mizuguchi Hiroyuki, Hayakawa Takao, Cattaneo Roberto, Russell Stephen J
Molecular Medicine Program, Mayo Foundation, 200 First St. SW, Rochester, Minnesota 55905, USA.
Nat Biotechnol. 2004 Mar;22(3):331-6. doi: 10.1038/nbt942. Epub 2004 Feb 15.
Membrane fusion has many potential applications in biotechnology. Here we show that antibody-targeted cell fusion can be achieved by engineering a fusogenic viral membrane glycoprotein complex. Three different single-chain antibodies were displayed at the extracellular C terminus of the measles hemagglutinin (H) protein, and combinations of point mutations were introduced to ablate its ability to trigger fusion through the native viral receptors CD46 and SLAM. When coexpressed with the measles fusion (F) protein, using plasmid cotransfection or bicistronic adenoviral vectors, the retargeted H proteins could mediate antibody-targeted cell fusion of receptor-negative or receptor-positive index cells with receptor-positive target cells. Adenoviral expression vectors mediating human epidermal growth factor receptor (EGFR)-targeted cell fusion were potently cytotoxic against EGFR-positive tumor cell lines and showed superior antitumor potency against EGFR-positive tumor xenografts as compared with control adenoviruses expressing native (untargeted) or CD38-targeted H proteins.
膜融合在生物技术领域有许多潜在应用。在此我们表明，通过改造一种促融合病毒膜糖蛋白复合物可实现抗体靶向的细胞融合。三种不同的单链抗体展示在麻疹血凝素（H）蛋白的细胞外C末端，并引入点突变组合以消除其通过天然病毒受体CD46和SLAM触发融合的能力。当与麻疹融合（F）蛋白共表达时，利用质粒共转染或双顺反子腺病毒载体，重新靶向的H蛋白可介导受体阴性或受体阳性指示细胞与受体阳性靶细胞的抗体靶向细胞融合。介导人表皮生长因子受体（EGFR）靶向细胞融合的腺病毒表达载体对EGFR阳性肿瘤细胞系具有强大的细胞毒性，并且与表达天然（非靶向）或CD38靶向H蛋白的对照腺病毒相比，对EGFR阳性肿瘤异种移植物显示出更强的抗肿瘤效力。
