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Reprogramming a Doxycycline-Inducible Gene Switch System for Bacteria-Mediated Cancer Therapy

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Abstract

Purpose

Attenuated Salmonella typhimurium is a potential biotherapeutic antitumor agent because it can colonize tumors and inhibit their growth. The present study aimed to develop a doxycycline (Doxy)-inducible gene switch system in attenuated S. typhimurium and assess its therapeutic efficacy in various tumor-bearing mice models.

Procedures

A Doxy-inducible gene switch system comprising two plasmids was engineered to trigger the expression of cargo genes (Rluc8 and clyA). Attenuated S. typhimurium carrying Rluc8 were injected intravenously into BALB/c mice bearing CT26 tumors, and bioluminescence images were captured at specified intervals post-administration of doxycycline. The tumor-suppressive effects of bacteria carrying clyA were evaluated in BALB/c mice bearing CT26 tumors and in C57BL/6 mice bearing MC38 tumors.

Results

Expression of the fimE gene, induced only in the presence of Doxy, triggered a unidirectional switch of the POXB20 promoter to induce expression of the cargo genes. The switch event was maintained over a long period of bacterial culture. After intravenous injection of transformed Salmonella into mice bearing CT26 tumors, the bacteria transformed with the Doxy-inducible gene switch system for Rluc8 targeted only tumor tissues and expressed the payloads 2 days after Doxy treatment. Notably, bacteria carrying the Doxy-inducible gene switch system for clyA effectively suppressed tumor growth and prolonged survival, even after just one Doxy induction.

Conclusions

These results suggest that attenuated S. typhimurium carrying this novel gene switch system elicited significant therapeutic effects through a single induction triggering and were a potential biotherapeutic agent for tumor therapy.

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Funding

This work was supported by the Bio & Medical Technology Development Program of National Research Foundation of Korea (NRF) (No. NRF-2020M3A9G3080282), NRF grants (Nos. 2020R1A5A2031185, 2021M3C1C3097637, 2021C300) funded by Ministry of Science and IT (MSIT), and by the Korea Drug Development Fund funded by Ministry of Science and ICT, Ministry of Trade, Industry, and Energy, and Ministry of Health and Welfare (HN22C0637, RS-2022-00167101). Y.H. was supported by the Bio & Medical Technology Development Program of NRF (No. NRF-2020M3A9G3080330).

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Authors and Affiliations

  1. Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea

    Hien Thi-Thu Ngo, Dinh-Huy Nguyen, Sung-Hwan You, Khuynh Van Nguyen, So-Young Kim & Jung-Joon Min

  2. Department of Molecular Medicine (BrainKorea21 Plus), Chonnam National University Graduate School, Gwangju, 61469, Republic of Korea

    Hien Thi-Thu Ngo, Dinh-Huy Nguyen, Khuynh Van Nguyen & Jung-Joon Min

  3. Department of Biochemistry, Hanoi Medical University, No 1, Ton That Tung St., Dong Da, Hanoi, 100000, Vietnam

    Hien Thi-Thu Ngo

  4. CNCure Biotech, Hwasun, 58128, Republic of Korea

    Sung-Hwan You, So-Young Kim, Yeongjin Hong & Jung-Joon Min

  5. Department of Microbiology, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea

    Yeongjin Hong

  6. Department of Nuclear Medicine, Chonnam National University Medical School and Hwasun Hospital, Gwangju, 61469, Republic of Korea

    Jung-Joon Min

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  1. Hien Thi-Thu Ngo
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Correspondence to Yeongjin Hong or Jung-Joon Min.

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Ngo, H.TT., Nguyen, DH., You, SH. et al. Reprogramming a Doxycycline-Inducible Gene Switch System for Bacteria-Mediated Cancer Therapy. Mol Imaging Biol 26, 148–161 (2024). https://doi.org/10.1007/s11307-023-01879-6

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  • DOI: https://doi.org/10.1007/s11307-023-01879-6

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