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The long persistence of pyrrolizidine alkaloid-derived DNA adducts in vivo: kinetic study following single and multiple exposures in male ICR mice

  • Genotoxicity and Carcinogenicity
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Abstract

Pyrrolizidine alkaloid (PA)-containing plants are widespread in the world and the most common poisonous plants affecting livestock, wildlife, and humans. Our previous studies demonstrated that PA-derived DNA adducts can potentially be a common biological biomarker of PA-induced liver tumor formation. In order to validate the use of these PA-derived DNA adducts as a biomarker, it is necessary to understand the basic kinetics of the PA-derived DNA adducts formed in vivo. In this study, we studied the dose-dependent response and kinetics of PA-derived DNA adduct formation and removal in male ICR mice orally administered with a single dose (40 mg/kg) or multiple doses (10 mg/kg/day) of retrorsine, a representative carcinogenic PA. In the single-dose exposure, the PA-derived DNA adducts exhibited dose-dependent linearity and persisted for up to 4 weeks. The removal of the adducts following a single-dose exposure to retrorsine was biphasic with half-lives of 9 h (t 1/2α) and 301 h (~12.5 days, t 1/2β). In the 8-week multiple exposure study, a marked accumulation of PA-derived DNA adducts without attaining a steady state was observed. The removal of adducts after the multiple exposure also demonstrated a biphasic pattern but with much extended half-lives of 176 h (~7.33 days, t 1/2α) and 1736 h (~72.3 days, t 1/2β). The lifetime of PA-derived DNA adducts was more than 8 weeks following the multiple-dose treatment. The significant persistence of PA-derived DNA adducts in vivo supports their role in serving as a biomarker of PA exposure.

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Abbreviations

PAs:

Pyrrolizidine alkaloids

dehydro-PAs:

Dehydropyrrolizidine alkaloids

DHP:

(±)-6,7-Dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine

dG:

2′-Deoxyguanosine

dA:

2′-Deoxyadenosine

DHP-dG-1 and DHP-dG-2:

A pair of epimers of 7-(deoxyguanosin-N 2-yl)dehydrosupinidine adducts

DHP-dG-3 and DHP-dG-4:

A pair of epimers of 7-hydroxy-9-(deoxyguanosin-N 2-yl)dehydrosupinidine adducts

DHP-dA-1 and DHP-dA-2:

A pair of epimers of 7-(deoxyadenosin-N 6-yl)dehydrosupinidine adducts

DHP-dA-3 and DHP-dA-4:

A pair of epimers of 7-hydroxy-9-(deoxyadenosin-N 6-yl)dehydrosupinidine adducts

HPLC–ES–MS/MS:

High-performance liquid chromatography-electrospray ionization tandem mass spectrometry

MRM:

Multiple reaction monitoring

LOD:

Limit of detection

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Acknowledgments

The present study was supported by Research Grant Council of Hong Kong (GRF Grant Nos. 471310 and 469712), CUHK Direct Grants (4054047 and 4054134), CUHK One-off Funding for Joint Lab/Research Collaboration (Project Code: 3132968), and CUHK School of Biomedical Sciences–Seed Fund for Joint Establishments. This article is not an official U.S. Food and Drug Administration (FDA) guidance or policy statement. No official support or endorsement by the U.S. FDA is intended or should be inferred.

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    Authors and Affiliations

    1. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China

      Lin Zhu, Junyi Xue & Ge Lin

    2. Joint Research Laboratory of Promoting Globalization of Traditional Chinese Medicines Between The Chinese University of Hong Kong and Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China

      Lin Zhu, Junyi Xue & Ge Lin

    3. National Center for Toxicological Research, Jefferson, AR, 72079, USA

      Qingsu Xia & Peter P. Fu

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    1. Lin Zhu
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    2. Junyi Xue
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    Lin Zhu and Junyi Xue have contributed equally to this work.

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    Zhu, L., Xue, J., Xia, Q. et al. The long persistence of pyrrolizidine alkaloid-derived DNA adducts in vivo: kinetic study following single and multiple exposures in male ICR mice. Arch Toxicol 91, 949–965 (2017). https://doi.org/10.1007/s00204-016-1713-z

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