Tumor and vascular targeting of a novel oncolytic measles virus retargeted against the urokinase receptor.
Tumor and vascular targeting of a novel oncolytic measles virus retargeted against the urokinase receptor.
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DOI：
10.1158/0008-5472.can-08-2628
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发表时间：
2009-02-15
期刊：
影响因子：
11.2
通讯作者：
Merchan JR
中科院分区：
文献类型：
--
作者：
Jing Y;Tong C;Zhang J;Nakamura T;Iankov I;Russell SJ;Merchan JR
溶瘤性麻疹病毒（MV）以CD46依赖的方式诱导细胞融合和细胞毒性。开发完全重靶向的溶瘤病毒将提高肿瘤的选择性。尿激酶受体（uPAR）是多种恶性肿瘤中过表达的肿瘤和基质靶点。将MV-H糖蛋白完全重靶向人或鼠uPAR，并测定其促聚变活性。对表达eGFP的重组人（MV-h-uPA）和鼠（MV-m-uPA） uPAR重组mv进行了营救和鉴定。RT-PCR和Western Blot检测嵌合MV-H的病毒表达。体外病毒复制与MV-GFP对照相当。在不同uPAR表达水平的人和小鼠细胞中证实了MV-uPAs的受体和物种特异性。通过Xa因子处理去除MV-uPA的ATF配体-削弱MV-uPA的功能活性。在表达uPAR的人和小鼠细胞中证实了细胞毒性。MV-h-uPA在体外可有效感染人内皮细胞和毛细血管。在MDA-MB-231乳腺癌异种移植模型中，静脉注射MV-h-uPA可延缓肿瘤生长并延长生存期。免疫组化证实病毒靶向肿瘤。肿瘤内给药后，MV-m-uPA在体内转导小鼠乳腺肿瘤（4T1）。MDA-MB-231模型的肿瘤毛细血管双（CD31和MV-N）染色证实，MV-m-uPA在全身给药后靶向小鼠肿瘤血管。总之，MV- upa是一种新型的溶瘤MV，具有强效和特异性的抗肿瘤作用和肿瘤血管靶向性。这是第一个能够在小鼠细胞中复制并以uPAR依赖的方式靶向肿瘤血管的重靶向溶瘤MV。
Oncolytic measles virus (MV) induces cell fusion and cytotoxicity in a CD46 dependent manner. Development of fully retargeted oncolytic MVs would improve tumor selectivity. The urokinase receptor (uPAR) is a tumor and stromal target overexpressed in multiple malignancies. MV-H glycoproteins fully retargeted to either human or murine uPAR were engineered and their fusogenic activity was determined. Recombinant human (MV-h-uPA) and murine (MV-m-uPA) uPAR retargeted MVs expressing eGFP were rescued and characterized. Viral expression of chimeric MV-H was demonstrated by RT-PCR and Western Blot. In vitro viral replication was comparable to MV-GFP control. Receptor and species specificity of MV-uPAs were demonstrated in human and murine cells with different levels of uPAR expression. Removal of the ATF ligand from MV-uPA -by Factor Xa treatment- ablated MV-uPA’s functional activity. Cytotoxicity was demonstrated in uPAR expressing human and murine cells. MV-h-uPA efficiently infected human endothelial cells and capillary tubes in vitro. Intravenous administration of MV-h-uPA delayed tumor growth and prolonged survival in the MDA-MB-231 breast cancer xenograft model. Viral tumor targeting was confirmed by immunohistochemistry. MV-m-uPA transduced murine mammary tumors (4T1) in vivo, after intratumor administration. MV-m-uPA targeted murine tumor vasculature after systemic administration, as demonstrated by dual (CD31 and MV-N) staining of tumor capillaries in the MDA-MB-231 model. In conclusion, MV-uPA is a novel oncolytic MV associated with potent and specific antitumor effects and tumor vascular targeting. This is the first retargeted oncolytic MV able to replicate in murine cells and target tumor vasculature in an uPAR dependent manner.