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Metabolomic Evaluation of N-Acetyl-p-Benzoquinone Imine Protein Adduct Formation with Therapeutic Acetaminophen Administration: Sex-based Physiologic Differences

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Abstract

Background

Acetaminophen (APAP)-associated transaminase elevation, induced by N-acetyl-p-benzoquinone imine (NAPQI) protein adduction, remains an area of research interest. Distinct from known genetic, physiologic, and dosage associations dictating severity of hepatic injury, no known factors predict an absence of protein adduct formation at therapeutic APAP dosing.

Hypothesis

Sex-based physiology is predictive of APAP-induced protein adduct formation and differential metabolite expression at therapeutic doses.

Methods

This retrospective study interrogated serum samples collected for a prior study investigating fluctuations of alanine aminotransferase (ALT) over time with 4G daily APAP dosing for ≥ 16 days in subjects from Denver, Colorado. Subjects were grouped by adduct formation (n = 184) vs no adducts (n = 20). Samples were run on ultra-high-performance liquid chromatography mass spectrometry from study days 0, 7, 16, and 31. Significant metabolite expressions were identified using t-tests with false discovery rate correction (FDR), partial least squares discriminant, and ANOVA simultaneous comparison analyses. Demographic and clinical data were explored using t-tests with FDR (age, weight, BMI, ALT) and Chi-square (sex, ethnicity, race) analyses.

Results

In pre-treatment samples, relative quantitation caprylic acid was expressed ninefold higher and 6-carboxyhexanoate was expressed threefold lower in subjects who did not develop adducts. Lactate had greater expression in the no adducts group (p = 0.001). Using absolute quantitation, glutathione was expressed 2.6-fold greater among no adduct subjects. Odds of males developing NAPQI protein adducts at therapeutic APAP dosing were 5.91 times lower than females (95% CI = 2.3–14.9; p = 0.0001).

Conclusion

Multiple metabolites were differentially expressed based on adduct group and sex. Metabolites were identified unique to adduct development independent of sex. At therapeutic APAP dosing, males were less likely to develop APAP protein adducts. Further research into lipid biosynthesis and metabolism may provide further insight into physiology associated with adduct production.

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Funding

The parent study was funded by Johnson and Johnson. A.A.M. received support from NIH R35GM124939 and NIH CTSI UL1 TR001082 for this work.

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Authors and Affiliations

  1. Department of Emergency Medicine, University of Colorado School of Medicine, University of Colorado Denver-Anschutz Medical Center, Mail Stop B-215, 12401 East 17thAvenue, Aurora, CO, 80045, USA

    Cosby G. Arnold, Layne Dylla, Andrew A. Monte, Kennon Heard & Brandon Sonn

  2. Rocky Mountain Poison & Drug Center, Denver Health and Hospital Authority, 1391 Speer Blvd Unit 600, Denver, CO, 80204, USA

    Andrew A. Monte, Kennon Heard, Susan Heard, Kate Reynolds, Richard Dart & Barry Rumack

  3. Division of Bioinformatics & Personalized Medicine, University of Colorado School of Medicine, 13001 E 17th Place, Fitzimons Building, 5th floor, #563, Aurora, CO, 80045, USA

    Angelo D’Alessandro

  4. Department of Pediatrics, University of Colorado School of Medicine, 13123 E 16th Ave, Aurora, CO, 80045, USA

    Barry Rumack

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  1. Cosby G. Arnold
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  2. Layne Dylla
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  7. Kate Reynolds
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  10. Brandon Sonn
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Corresponding author

Correspondence to Cosby G. Arnold.

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Conflict of Interest

None. However, the original project that collected the samples used for this study was supported by an investigator-initiated grant from McNeil Consumer Healthcare. Authors received only salary for their work on the project.

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Supervising Editor: Anselm Wong, MBBS DipTox PhD

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Prior Presentations: Data in this manuscript were previously presented as a platform at ACMT’s Annual Scientific Meeting, Virtual, 2021.

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Arnold, C.G., Dylla, L., Monte, A. et al. Metabolomic Evaluation of N-Acetyl-p-Benzoquinone Imine Protein Adduct Formation with Therapeutic Acetaminophen Administration: Sex-based Physiologic Differences. J. Med. Toxicol. 18, 297–310 (2022). https://doi.org/10.1007/s13181-022-00903-5

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  • DOI: https://doi.org/10.1007/s13181-022-00903-5

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