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The tunicamycin derivative TunR2 exhibits potent antibiotic properties with low toxicity in an in vivo Mycobacterium marinum-zebrafish TB infection model

The Journal of Antibiotics volume 77, pages 245–256 (2024)Cite this article

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Abstract

Tunicamycins (TUN) are well-defined, Streptomyces-derived natural products that inhibit protein N-glycosylation in eukaryotes, and by a conserved mechanism also block bacterial cell wall biosynthesis. TUN inhibits the polyprenylphosphate-N-acetyl-hexosamine-1-phospho-transferases (PNPT), an essential family of enzymes found in both bacteria and eukaryotes. We have previously published the development of chemically modified TUN, called TunR1 and TunR2, that have considerably reduced activity on eukaryotes but that retain the potent antibacterial properties. A mechanism for this reduced toxicity has also been reported. TunR1 and TunR2 have been tested against mammalian cell lines in culture and against live insect cells but, until now, no in vivo evaluation has been undertaken for vertebrates. In the current work, TUN, TunR1, and TunR2 are investigated for their relative toxicity and antimycobacterial activity in zebrafish using a well-established Mycobacterium marinum (M. marinum) infection system, a model for studying human Mycobacterium tuberculosis infections. We also report the relative ability to activate the unfolded protein response (UPR), the known mechanism for the eukaryotic toxicity observed with TUN treatment. Importantly, TunR1 and TunR2 retained their antimicrobial properties, as evidenced by a reduction in M. marinum bacterial burden, compared to DMSO-treated zebrafish. In summary, findings from this study highlight the characteristics of recently developed TUN derivatives, mainly TunR2, and its potential for use as a novel anti-bacterial agent for veterinary and potential medical purposes.

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Acknowledgements

This work was supported in part by the U.S. Department of Agriculture, Agricultural Research Service (N.P.J.P. and M.A.J.), and by Eminent Scholar funds from the Medical College of Wisconsin (B.A.L). T.C.Z. was supported by the National Institute of Allergy and Infectious Diseases (5R21AI144225-02). We acknowledge C. Skory (NCAUR-ARS, Peoria, IL) and J. Bannantine (NADC-ARS, Ames, IA) for helpful discussion and suggestions on the preparation of this manuscript. We thank T. Hartman for expert technical support and David M. Tobin (Duke University) for sharing reagents. Finally we acknowledge the use of BioRender for the generation of diagrams used in this manuscript. Mention of trade names or commercial products in this article is solely for the purpose of providing scientific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.

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Authors and Affiliations

  1. Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI, USA

    Hannah J. T. Nonarath & Brian A. Link

  2. USDA, Agricultural Research Service, Renewable Products Technology Research, National Center for Agricultural Utilization Research, 1815 N. University St., Peoria, IL, 61604, USA

    Michael A. Jackson & Neil P. J. Price

  3. Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, USA

    Renee M. Penoske & Thomas C. Zahrt

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Nonarath, H.J.T., Jackson, M.A., Penoske, R.M. et al. The tunicamycin derivative TunR2 exhibits potent antibiotic properties with low toxicity in an in vivo Mycobacterium marinum-zebrafish TB infection model. J Antibiot 77, 245–256 (2024). https://doi.org/10.1038/s41429-023-00694-z

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