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Published on: August 1, 2019
Rescue of NanoLuc luciferase-expressing Senecavirus A with oncolytic activity
Fuxiao Liu1, Qianqian Wang1, Yilan Huang1
1College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, 266109, China.
Abstract:
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.
Insights
Senecavirus A (SVA) engineered with NanoLuc® luciferase (NLuc) shows oncolytic potential against small cell lung cancer cells. This recombinant virus (rSVA-NLuc) enables efficient imaging for oncolytic virotherapy studies.
Area of Science:
- Virology
- Oncolytic Virotherapy
- Molecular Biology
Background:
- Senecavirus A (SVA) is a picornavirus with oncolytic activity against neuroendocrine tumor cells, including small cell lung cancer (SCLC).
- Developing effective imaging tools is crucial for assessing oncolytic virotherapy efficacy in vivo.
Purpose of the Study:
- To rescue and characterize a recombinant SVA expressing NanoLuc® luciferase (NLuc) for oncolytic analysis.
- To evaluate the utility of NLuc-tagged SVA (rSVA-NLuc) in monitoring viral replication and oncolysis in SCLC cells.
Main Methods:
- Rescue of a recombinant SVA encoding NLuc.
- Characterization of viral replication and NLuc expression kinetics in NCI-H446 SCLC cells.
- Assessment of the oncolytic phenotype induced by rSVA-NLuc.
Main Results:
- rSVA-NLuc demonstrated efficient viral replication and rapid NLuc expression in H446 cells.
- Infected H446 cells displayed an oncolytic phenotype (rounding, detachment, lysis) within 48 hours.
- NLuc expression facilitated real-time monitoring of viral activity and oncolysis.
Conclusions:
- The NLuc reporter system is effective for analyzing the oncolytic activity of SVA in vitro.
- rSVA-NLuc holds potential for in vivo imaging applications in oncolytic virotherapy due to NLuc's advantages for scalability.
