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Engineered Mycobacterium tuberculosis triple-kill-switch strain provides controlled tuberculosis infection in animal models

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Title: Engineered Mycobacterium tuberculosis triple-kill-switch strain provides controlled tuberculosis infection in animal models
Authors: Xin Wang, Hongwei Su, Joshua B. Wallach, Jeffrey C. Wagner, Benjamin J. Braunecker, Michelle Gardner, Kristine M. Guinn, Nicole C. Howard, Thais Klevorn, Kan Lin, Yue J. Liu, Yao Liu, Douaa Mugahid, Mark Rodgers, Jaimie Sixsmith, Shoko Wakabayashi, Junhao Zhu, Matthew Zimmerman, Véronique Dartois, JoAnne L. Flynn, Philana Ling Lin, Sabine Ehrt, Sarah M. Fortune, Eric J. Rubin, Dirk Schnappinger
Source: Nat Microbiol
Publisher Information: Springer Science and Business Media LLC, 2025.
Publication Year: 2025
Subject Terms: Trimethoprim/pharmacology, Animal, Mycobacteriophages/genetics/physiology, Bacterial Proteins/genetics/metabolismAntitubercular Agents/pharmacology, Antitubercular Agents, Mycobacterium tuberculosis, Mice, SCID, Mycobacteriophages, SCID, Article, Mice, Disease Models, Animal, Bacterial Proteins, Disease Models, Operon, Animals, Tuberculosis, Humans, Tuberculosis/microbiology, Female, Genetic Engineering, WCM Library Coordinated Deposit
Description: Human challenge experiments could accelerate tuberculosis vaccine development. This requires a safe Mycobacterium tuberculosis (Mtb) strain that can both replicate in the host and be reliably cleared. Here we genetically engineered Mtb strains encoding up to three kill switches: two mycobacteriophage lysin operons negatively regulated by tetracycline and a degron domain–NadE fusion, which induces ClpC1-dependent degradation of the essential enzyme NadE, negatively regulated by trimethoprim. The triple-kill-switch (TKS) strain showed similar growth kinetics and antibiotic susceptibilities to wild-type Mtb under permissive conditions but was rapidly killed in vitro without trimethoprim and doxycycline. It established infection in mice receiving antibiotics but was rapidly cleared upon cessation of treatment, and no relapse was observed in infected severe combined immunodeficiency mice or Rag−/− mice. The TKS strain had an escape mutation rate of less than 10−10 per genome per generation. These findings suggest that the TKS strain could be a safe, effective candidate for a human challenge model.
Document Type: Article
Other literature type
File Description: application/pdf
Language: English
ISSN: 2058-5276
DOI: 10.1038/s41564-024-01913-5
Access URL: https://pubmed.ncbi.nlm.nih.gov/39794471
Rights: CC BY
CC BY NC ND
Accession Number: edsair.doi.dedup.....f880d58b6a31e7eef5b81e65c723a7f9
Database: OpenAIRE
Description
Abstract:Human challenge experiments could accelerate tuberculosis vaccine development. This requires a safe Mycobacterium tuberculosis (Mtb) strain that can both replicate in the host and be reliably cleared. Here we genetically engineered Mtb strains encoding up to three kill switches: two mycobacteriophage lysin operons negatively regulated by tetracycline and a degron domain–NadE fusion, which induces ClpC1-dependent degradation of the essential enzyme NadE, negatively regulated by trimethoprim. The triple-kill-switch (TKS) strain showed similar growth kinetics and antibiotic susceptibilities to wild-type Mtb under permissive conditions but was rapidly killed in vitro without trimethoprim and doxycycline. It established infection in mice receiving antibiotics but was rapidly cleared upon cessation of treatment, and no relapse was observed in infected severe combined immunodeficiency mice or Rag−/− mice. The TKS strain had an escape mutation rate of less than 10−10 per genome per generation. These findings suggest that the TKS strain could be a safe, effective candidate for a human challenge model.
ISSN:20585276
DOI:10.1038/s41564-024-01913-5