Tumor-targeting bioluminescent bacteria for in vivo imaging

Abstract

As the understanding of bacteria-mediated cancer therapies deepens, bacteria such as Escherichia coli Nissle 1917 (EcN) have become a promising platform for cancer therapy. However, their potential role in real-time monitoring and visualization tools still needs to be explored and enhanced. In this study, we aimed to screen and optimize EcN visualization systems for non-invasive in vivo bioluminescence imaging in live mice. To this end, we developed three series of recombinant EcN strains expressing Gaussia luciferase (Gluc), Renilla luciferase (Rluc), and NanoLuc (Nluc), along with their respective mutants. These strains exhibited bioluminescence when different coelenterazine (CTZ) substrates were present. As a result, multiple bioluminescent EcN strain-substrate pairs were identified with stronger, longer, or red-shifted bioluminescence, offering multiple effective optical tumor-targeting systems for in vivo studies investigating bacteria-mediated cancer therapy and intestinal diseases.

Keywords

Escherichia coli Nissle 1917 / In vivo bioluminescence imaging / Luciferase / Coelenterazine

Data Availability Statement

All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials.

Declaration of Competing Interest

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Given his role as Editor-in-Chief, Dr. Youming Zhang, had no involvement in the peer-review of this article and has no access to information regarding its peer-review. Full responsibility for the editorial process for this article was delegated to Dr. Dongchun Ni.

CRediT authorship contribution statement

Chenghao Ma: Writing - original draft, Methodology, Data curation. Jingxi Liu: Methodology. Hongjing Liu: Methodology, Data curation. Xiaohan Zhao: Methodology, Data curation. Geng Li: Methodology. Youming Zhang: Writing - review & editing, Supervision. Tianyu Jiang: Writing - review & editing, Supervision, Conceptualization.

Acknowledgments

Thanks to Xiaomin Zhao, Haiyan Yu, Changbin Liu, Yuyu Guo and Sen Wang from the Core Facilities for Life and Environmental Sciences at the SKLMT (State Key Laboratory of Microbial Technology, Shandong University) for the assistance provided in in vivo imaging. Thanks to Prof. Minyong Li and Dr. Xingye Yang from Key Laboratory of Chemical Biology (MOE), Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University for the supply of CTZ analogues including CTZ, CTZ B5, CTZ B12, CTZ h, and FRZ.

This study was supported by the National Natural Science Foundation of China (No. 32201245 and No. 32170038), the Shandong Provincial Natural Science Foundation (ZR2020QB158), the Guangdong Basic and Applied Basic Research Foundation (2020A1515110284), the Major International Joint Research Project of the National Natural Science Foundation of China (No. 32161133013), the Future Plan for Young Scholars of Shandong University, and the SKLMT Frontiers and Challenges Project SKLMTFCP-2023-05.

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