Senecavirus A (SVA), previously known as Seneca Valley virus, is classified into the genus Senecavirus in the family Picornaviridae. SVA is not pathogenic to normal human cells, but has potent oncolytic activity in some tumor cells with neuroendocrine feature, such as small cell lung cancer (SCLC) NCI-H446 cell line. In this study, we rescued and characterized a recombinant SVA that could efficiently express a novel luciferase, NanoLuc® luciferase (NLuc), which was smaller and “brighter” than others. This NLuc-tagged recombinant SVA (rSVA-NLuc) exhibited high capacity for viral replication, but genetic instability of NLuc during serial virus passages. The NLuc as a reporter facilitated oncolytic analysis of rSVA-NLuc in H446 cells. The rSVA-NLuc-infected H446 cells exhibited an oncolytic phenotype characterized by cell rounding, swelling, detachment and lysis at 48 h post infection. Kinetic curve showed that the NLuc was rapidly expressed in H446 cells during an exponential phase of viral growth. Because the NLuc offered several advantages over fluorescent proteins for assay scalability in vivo, the rSVA-NLuc would play a potential role in facilitating in vivo imaging studies of oncolytic virotherapy.

Senecavirus A（SVA），以前称为塞内卡谷病毒，分为属Senecavirus在家庭中的小核糖核酸病毒. SVA 对正常人类细胞没有致病性，但在一些具有神经内分泌特征的肿瘤细胞中具有有效的溶瘤活性，例如小细胞肺癌 (SCLC) NCI-H446 细胞系。在这项研究中，我们拯救并表征了一种重组 SVA，它可以有效表达一种新型荧光素酶 NanoLuc® 荧光素酶 (NLuc)，它比其他荧光素酶更小、更“亮”。这种带有 NLuc 标签的重组 SVA (rSVA-NLuc) 表现出很高的病毒复制能力，但在连续病毒传代过程中 NLuc 的遗传不稳定性。NLuc 作为报告基因促进了 H446 细胞中 rSVA-NLuc 的溶瘤分析。rSVA-NLuc 感染的 H446 细胞表现出溶瘤表型，其特征是在感染后 48 小时时细胞变圆、肿胀、脱离和裂解。动力学曲线显示 NLuc 在病毒生长的指数期期间在 H446 细胞中快速表达。因为 NLuc 在测定可扩展性方面提供了优于荧光蛋白的几个优势在体内，rSVA-NLuc 将在促进溶瘤病毒疗法的体内成像研究方面发挥潜在作用。