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Pro-caspase-3 protects cells from polymyxin B-induced cytotoxicity by preventing ROS accumulation


Polymyxin B (PMB), a last-line antibiotic used against antibiotic-resistant superbugs, causes undesirable cytotoxic side effects. However, its mechanisms remain unknown. In this study, we unexpectedly found that caspase-3, a main executor of apoptosis, plays a protective role in PMB-induced cytotoxicity. Caspase-3 knockout (KO) cells exhibited higher susceptibility to PMB-induced cytotoxicity compared with wild-type (WT) cells, accompanied by increased levels of reactive oxygen species (ROS). Interestingly, co-treatment with the antioxidant N-acetylcysteine (NAC) rescued cell viability to a similar extent as WT cells. Furthermore, PMB failed to facilitate the processing of inactive caspase-3 (pro-caspase-3) into active forms, suggesting that pro-caspase-3 nonenzymatically suppresses PMB-driven ROS accumulation and its cytotoxicity. Thus, our findings that demonstrate the potential ability of PMB to stimulate ROS generation, but which is normally masked by pro-caspase-3-dependent mechanisms, may provide novel insights into the mechanisms of PMB-induced side effects.

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  1. 1.

    Nord NM, Hoeprich PD, Polymyxin B, Colistin. A. Critical Comparison. New Engl J Med. 1964;270:1030–5.

    CAS  Article  Google Scholar 

  2. 2.

    Arnold TM, Forrest GN, Messmer KJ. Polymyxin antibiotics for gram-negative infections. Am J Health-Syst Pharm. 2007;64:819–26.

    CAS  Article  Google Scholar 

  3. 3.

    Khondker A, Dhaliwal AK, Saem S, Mahmood A, Fradin C, Moran-Mirabal J, et al. Membrane charge and lipid packing determine polymyxin-induced membrane damage. Commun Biol. 2019;2:67.

    Article  Google Scholar 

  4. 4.

    Velkov T, Thompson PE, Nation RL, Li J. Structure–activity relationships of polymyxin antibiotics. J Med Chem. 2010;53:1898–916.

    CAS  Article  Google Scholar 

  5. 5.

    Li J, Nation RL. Old polymyxins are back: is resistance close? Clin Infect Dis. 2006;43:663–4.

    Article  Google Scholar 

  6. 6.

    Cai Y, Lee W, Kwa AL. Polymyxin B versus colistin: an update. Expert Rev anti-Infect Ther. 2015;13:1481–97.

    CAS  Article  Google Scholar 

  7. 7.

    Kubin CJ, Ellman TM, Phadke V, Haynes LJ, Calfee DP, Yin MT. Incidence and predictors of acute kidney injury associated with intravenous polymyxin B therapy. J Infect. 2012;65:80–87.

    Article  Google Scholar 

  8. 8.

    Azad MA, Finnin BA, Poudyal A, Davis K, Li J, Hill PA, et al. Polymyxin B induces apoptosis in kidney proximal tubular cells. Antimicrob agents Chemother. 2013;57:4329–35.

    CAS  Article  Google Scholar 

  9. 9.

    Noguchi T, Tsuchida M, Kogue Y, Spadini C, Hirata Y, Matsuzawa A. Brefeldin A-inhibited guanine nucleotide-exchange factor 1 (BIG1) governs the recruitment of tumor necrosis factor receptor-associated factor 2 (TRAF2) to tumor necrosis factor receptor 1 (TNFR1) signaling complexes. Int J Mol Sci. 2016;17:1869.

    Article  Google Scholar 

  10. 10.

    Hirata Y, Katagiri K, Nagaoka K, Morishita T, Kudoh Y, Hatta T, et al. TRIM48 Promotes ASK1 activation and cell death through ubiquitination-dependent degradation of the ASK1-negative regulator PRMT1. Cell Rep. 2017;21:2447–57.

    CAS  Article  Google Scholar 

  11. 11.

    Sekiguchi Y, Yamada M, Noguchi T, Noomote C, Tsuchida M, Kudoh Y, et al. The anti-cancer drug gefitinib accelerates Fas-mediated apoptosis by enhancing caspase-8 activation in cancer cells. J Toxicol Sci. 2019;44:435–40.

    Article  Google Scholar 

  12. 12.

    McArthur K, Kile BT. Apoptotic caspases: multiple or mistaken identities? Trends cell Biol. 2018;28:475–93.

    CAS  Article  Google Scholar 

  13. 13.

    Noguchi T, Suzuki M, Mutoh N, Hirata Y, Tsuchida M, Miyagawa S, et al. Nuclear-accumulated SQSTM1/p62-based ALIS act as microdomains sensing cellular stresses and triggering oxidative stress-induced parthanatos. Cell death Dis. 2018;9:1193.

    Article  Google Scholar 

  14. 14.

    Kim JS, Ha JY, Yang SJ, Son JH. A novel non-apoptotic role of procaspase-3 in the regulation of mitochondrial biogenesis activators. J Cell Biochem. 2018;119:347–57.

    CAS  Article  Google Scholar 

  15. 15.

    Brentnall M, Weir DB, Rongvaux A, Marcus AI, Boise LH. Procaspase-3 regulates fibronectin secretion and influences adhesion, migration and survival independently of catalytic function. J cell Sci. 2014;127:2217–26.

    CAS  Article  Google Scholar 

  16. 16.

    Dezoti Fonseca C, Watanabe M, Vattimo Mde F. Role of heme oxygenase-1 in polymyxin B-induced nephrotoxicity in rats. Antimicrob Agents Chemother. 2012;56:5082–7.

    Article  Google Scholar 

  17. 17.

    Suzuki T, Yamamoto M. Molecular basis of the Keap1-Nrf2 system. Free Radic Biol Med. 2015;88:93–100.

    CAS  Article  Google Scholar 

  18. 18.

    Chen M, Ona VO, Li M, Ferrante RJ, Fink KB, Zhu S, et al. Minocycline inhibits caspase-1 and caspase-3 expression and delays mortality in a transgenic mouse model of Huntington disease. Nat Med. 2000;6:797–801.

    CAS  Article  Google Scholar 

  19. 19.

    Krajewska M, Wang HG, Krajewski S, Zapata JM, Shabaik A, Gascoyne R, et al. Immunohistochemical analysis of in vivo patterns of expression of CPP32 (caspase-3), a cell death protease. Cancer Res. 1997;57:1605–13.

    CAS  PubMed  Google Scholar 

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This work was supported by JSPS KAKENHI Grant Numbers JP18H02567 and JP18K06622, and by MEXT KAKENHI JP17H05518 and JP19H05282. This work was also supported by the Fugaku Trust for Medicinal Research, the Takeda Science Foundation, and the Division for Interdisciplinary Advanced Research and Education (DIARE) Tohoku University.

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Correspondence to Takuya Noguchi or Atsushi Matsuzawa.

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Yokosawa, T., Yamada, M., Noguchi, T. et al. Pro-caspase-3 protects cells from polymyxin B-induced cytotoxicity by preventing ROS accumulation. J Antibiot 72, 848–852 (2019).

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