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Succinate prodrugs as treatment for acute metabolic crisis during fluoroacetate intoxication in the rat

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Abstract

Sodium fluoroacetate (FA) is a metabolic poison that systemically inhibits the tricarboxylic acid (TCA) cycle, causing energy deficiency and ultimately multi-organ failure. It poses a significant threat to society because of its high toxicity, potential use as a chemical weapon and lack of effective antidotal therapy. In this study, we investigated cell-permeable succinate prodrugs as potential treatment for acute FA intoxication. We hypothesized that succinate prodrugs would bypass FA-induced mitochondrial dysfunction, provide metabolic support, and prevent metabolic crisis during acute FA intoxication. To test this hypothesis, rats were exposed to FA (0.75 mg/kg) and treated with the succinate prodrug candidate NV354. Treatment efficacy was evaluated based on cardiac and cerebral mitochondrial respiration, mitochondrial content, metabolic profiles and tissue pathology. In the heart, FA increased concentrations of the TCA metabolite citrate (+ 4.2-fold, p < 0.01) and lowered ATP levels (− 1.9-fold, p < 0.001), confirming the inhibition of the TCA cycle by FA. High-resolution respirometry of cardiac mitochondria further revealed an impairment of mitochondrial complex V (CV)-linked metabolism, as evident by a reduced phosphorylation system control ratio (− 41%, p < 0.05). The inhibition of CV-linked metabolism is a novel mechanism of FA cardiac toxicity, which has implications for drug development and which NV354 was unable to counteract at the given dose. In the brain, FA induced the accumulation of β-hydroxybutyrate (+ 1.4-fold, p < 0.05) and the reduction of mitochondrial complex I (CI)-linked oxidative phosphorylation (OXPHOSCI) (− 20%, p < 0.01), the latter of which was successfully alleviated by NV354. This promising effect of NV354 warrants further investigations to determine its potential neuroprotective effects.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Abliva AB for providing NV354 and Dr. Christopher Petucci and the University of Pennsylvania Metabolomics Core for performing the metabolomics analysis. We would like to thank Drs. David Jett and Shardell Spriggs for their encouragement and guidance and the many members of the CounterAct Program who work tirelessly for our sake.

Funding

This work is supported by the CounterAct Program, National Institute of Neurologic Disorders and Stroke [Kilbaugh; R21 NS103826]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Resuscitation Science Center of Emphasis, The Children’s Hospital of Philadelphia, 3615 Civic Center Blvd, Philadelphia, PA, 19104, USA

    Sarah Piel, Joanna I. Janowska, J. Laurenson Ward, Meagan J. McManus, Danielle I. Aronowitz, Piotr K. Janowski, Jonathan Starr, Carly L. Clayman & Todd J. Kilbaugh

  2. Anesthesiology and Critical Care Medicine, The Children’s Hospital of Philadelphia, Philadelphia, USA

    Sarah Piel, Joanna I. Janowska, J. Laurenson Ward, Meagan J. McManus, Danielle I. Aronowitz, Piotr K. Janowski, Jonathan Starr, Carly L. Clayman & Todd J. Kilbaugh

  3. Department of Pathology, University of Iowa Carver College of Medicine, Iowa City, USA

    Jordan N. Hook & Marco M. Hefti

  4. Abliva AB, Lund, Sweden

    Eskil Elmér & Magnus J. Hansson

  5. Mitochondrial Medicine, Department of Clinical Sciences Lund, Lund University, Lund, Sweden

    Eskil Elmér, Magnus J. Hansson & Johannes K. Ehinger

  6. Department of Emergency Medicine, Division of Medical Toxicology, University of Pennsylvania School of Medicine, Philadelphia, USA

    David H. Jang

  7. Department of Neurosurgery, Rigshospitalet, Copenhagen, Denmark

    Michael Karlsson

  8. Otorhinolaryngology, Head and Neck Surgery, Department of Clinical Sciences Lund, Lund University, Skåne University Hospital, Lund, Sweden

    Johannes K. Ehinger

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  1. Sarah Piel
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Contributions

TJK, SP, JIJ, MK, and JKE contributed to the study conception and design. Material preparation, data collection, and analysis were performed by SP, JIJ, JLW, MH, EE, PKJ, JS, and JNH. The first draft of the manuscript was written by SP, JIJ, and JLW and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Sarah Piel.

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Competing interests

S.P., M.K., J.K.E., E.E. and M.J.H have, or have had, salary from and/or equity interest in Abliva AB. S.P., J.K.E., E.E. and M.J.H have filed patent applications for the use of succinate prodrugs for treatment of lactic acidosis or drug-induced side-effects due to complex I-related impairment of mitochondrial oxidative phosphorylation (WO/2015/155238). S.P., M.K., J.K.E., E.E. and M.J.H additionally have filed patent applications for the use of protected carboxylic acid-based metabolites for treatment of mitochondrial disorders (WO/2017/060400, WO/2017/060418, WO/2017/060422).

Ethics approval

All procedures were approved by the Institutional Animal Care and Use Committee at the Children’s Hospital of Philadelphia and performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Piel, S., Janowska, J.I., Ward, J.L. et al. Succinate prodrugs as treatment for acute metabolic crisis during fluoroacetate intoxication in the rat. Mol Cell Biochem 478, 1231–1244 (2023). https://doi.org/10.1007/s11010-022-04589-9

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