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Method | Open Access

Optimizing a high-sensitivity NanoLuc-based bioluminescence system for in vivo evaluation of antimicrobial treatment

Weilong Shang1,#,, Zhen Hu1,#, Mengyang Li2,#, Yuting Wang1, Yifan Rao3, Li Tan1, Juan Chen4, Xiaonan Huang1, Lu Liu2, He Liu1, Zuwen Guo1, Huagang Peng1, Yi Yang1, Qiwen Hu1, Shu Li1, Xiaomei Hu1( ), Jiao Zou5( ), Xiancai Rao1( )
Department of Microbiology, College of Basic Medical Sciences, Key Laboratory of Microbial Engineering Under the Educational Committee in Chongqing, Army Medical University (Third Military Medical University), Chongqing, China
Department of Microbiology, School of Medicine, Chongqing University, Chongqing, China
Department of Emergency Medicine, Xinqiao Hospital, Army Medical University (Third Military Medical University), Chongqing, China
Department of Pharmacy, Xinqiao Hospital, Army Medical University (Third Military Medical University), Chongqing, China
Department of Military Cognitive Psychology, School of Psychology, Army Medical University (Third Military Medical University), Chongqing, China

#These authors contributed equally to this work.

Edited by Wei Huang, University of Oxford, UK

Show Author Information

Abstract

Focal and systemic infections are serious threats to human health. Preclinical models enable the development of new drugs and therapeutic regimens. In vivo, animal bioluminescence (BL) imaging has been used with bacterial reporter strains to evaluate antimicrobial treatment effects. However, high-sensitivity bioluminescent systems are required because of the limited tissue penetration and low brightness of the BL signals of existing approaches. Here, we report that NanoLuc (Nluc) showed better performance than LuxCDABE in bacteria. However, the retention rate of plasmid constructs in bacteria was low. To construct stable Staphylococcus aureus reporter strains, a partner protein enolase (Eno) was identified by screening of S. aureus strain USA300 for fusion expression of Nluc-based luciferases, including Nluc, Teluc, and Antares2. Different substrates, such as hydrofurimazine (HFZ), furimazine (FUR), and diphenylterazine (DTZ), were used to optimize a stable reporter strain/substrate pair for BL imaging. S. aureus USA300/Eno-Antares2/HFZ produced the highest number of photons of orange-red light in vitro and enabled sensitive BL tracking of S. aureus in vivo, with sensitivities of approximately 10 CFU from mouse skin and 750 CFU from mouse kidneys. USA300/Eno-Antares2/HFZ was a powerful combination based on the longitudinal evaluation of the therapeutic efficacy of antibiotics. The optimized S. aureus Eno-Antares2/HFZ pair provides a technological advancement for the in vivo evaluation of antimicrobial treatment.

Keywords

Staphylococcus aureustherapeutic efficacybioluminescence imagingantimicrobial evaluationNluc-based luciferases

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mLife
Volume 2 Issue 4,
December 2023
Pages 462-478
DOI: 10.1002/mlf2.12091
Cite this article:
Shang W, Hu Z, Li M, et al. Optimizing a high-sensitivity NanoLuc-based bioluminescence system for in vivo evaluation of antimicrobial treatment. mLife, 2023, 2(4): 462-478. https://doi.org/10.1002/mlf2.12091

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Received: 01 November 2022
Accepted: 28 August 2023
Published: 20 December 2023
© 2023 The Authors. mLife published by John Wiley & Sons Australia, Ltd on behalf of Institute of Microbiology, Chinese Academy of Sciences.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
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