A Novel Oncolytic Chimeric Orthopoxvirus Encoding Luciferase Enables Real-Time View of Colorectal Cancer Cell Infection
A Novel Oncolytic Chimeric Orthopoxvirus Encoding Luciferase Enables Real-Time View of Colorectal Cancer Cell Infection
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DOI：
10.1016/j.omto.2018.03.001
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发表时间：
2018-06-29
期刊：
影响因子：
--
通讯作者：
Chen, Nanhai G.
中科院分区：
文献类型：
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作者：
O'Leary, Michael P.;Warner, Susanne G.;Chen, Nanhai G.
本研究假设一种新型溶瘤嵌合正痘病毒 CF33-Fluc 可成像并靶向结直肠癌细胞 (CRC)。构建了一种新型嵌合正痘病毒（CF33）。将胸苷激酶位点替换为萤火虫荧光素酶（Fluc），产生重组病毒-CF33-Fluc。进行了体外细胞毒性和病毒复制测定。体内 CRC 侧翼异种移植物接受单剂量瘤内或静脉内 CF33-Fluc。通过荧光素酶成像和器官滴度分析病毒生物分布。 CF33-Fluc 在体外以剂量依赖性方式感染、复制并杀死 CRC。与除一种亲本毒株外的所有毒株相比，CF33 具有更好的胞外包膜病毒分泌能力。无论 HCT-116 结直肠肿瘤异种移植模型中的病毒递送方法如何，低剂量在短时间内体内肿瘤都会快速消退或稳定。病毒感染的肿瘤细胞中荧光素酶的快速表达与治疗反应相关。 CRC 死亡通过坏死性凋亡途径发生。无论采用何种递送方式，CF33-Fluc 都会在体外和体内复制并杀死结直肠癌细胞。荧光素酶的表达能够实时跟踪病毒复制。尽管存在嵌合现象，CRC 死亡还是通过标准痘病毒诱导机制发生的。需要在免疫活性模型中进行进一步的研究。
This study hypothesizes that a novel oncolytic chimeric orthopoxvirus CF33-Fluc is imageable and targets colorectal cancer cells (CRCs). A novel chimeric orthopoxvirus (CF33) was constructed. The thymidine kinase locus was replaced with firefly luciferase (Fluc) to yield a recombinant virus-CF33-Fluc. In vitro cytotoxicity and viral replication assays were performed. In vivo CRC flank xenografts received single doses of intratumoral or intravenous CF33-Fluc. Viral biodistribution was analyzed via luciferase imaging and organ titers. CF33-Fluc infects, replicates in, and kills CRCs in vitro in a dose-dependent manner. CF33 has superior secretion of extracellular-enveloped virus versus all but one parental strain. Rapid tumor regression or stabilization occurred in vivo at a low dose over a short time period, regardless of the viral delivery method in the HCT-116 colorectal tumor xenograft model. Rapid luciferase expression in virus-infected tumor cells was associated with treatment response. CRC death occurs via necroptotic pathways. CF33-Fluc replicates in and kills colorectal cancer cells in vitro and in vivo regardless of delivery method. Expression of luciferase enables real-time tracking of viral replication. Despite the chimerism, CRC death occurs via standard poxvirus-induced mechanisms. Further studies are warranted in immunocompetent models.