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Mechanisms of CYP3A Induction During Pregnancy: Studies in HepaRG Cells

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Abstract

Activity of CYP3A, an enzyme responsible for metabolism of many marketed drugs, is induced by ~ 2-fold in pregnant women. Through studies in sandwich-cultured human hepatocytes (SCHH) and HepaRG cells, our laboratory has shown that this induction is likely mediated by the increase in cortisol plasma concentrations during pregnancy. Cortisol, at plasma concentrations observed during the third trimester (~ 800 nM), either alone or in combination with other pregnancy-related hormones, induces CYP3A activity in SCHH and HepaRG cells when cultured in dexamethasone-free media. To determine the mechanism(s) by which cortisol induces CYP3A activity, HepaRG cells were pre-incubated in dexamethasone-free medium and then incubated for 72 h with cortisol (798 nM). Glucocorticoid receptor (GR), pregnane X receptor (PXR), and CYP3A4 or CYP3A5 were knocked down using siRNA, and mRNA expression of these genes was measured. CYP3A4, and not CYP3A5, was found to be the dominant contributor to total CYP3A activity in control- and cortisol-treated HepaRG cells. Constitutive mRNA expression of CYP3A4 in HepaRG cells was regulated by both PXR and GR whereas constitutive expression of CYP3A5 in HepaRG cells was regulated by GR alone. Cortisol-mediated CYP3A4 induction in HepaRG cells was primarily mediated by GR-dependent PXR induction pathway and to a smaller extent via a PXR-independent pathway. Cortisol-mediated CYP3A5 induction was regulated by GR-dependent PXR-independent pathway. These data indicate that PXR plays a central role in cortisol-mediated induction of CYP3A activity during pregnancy and suggests that other enzymes and transporters, such as CYP2B6 and P-glycoprotein, may also be induced during pregnancy via the same mechanism(s).

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Abbreviations

SCHH:

Sandwich-cultured human hepatocytes

GR:

Glucocorticoid receptor

PXR:

Pregnane X receptor

UGT1A1:

UDP-glucuronosyltransferase 1A1

TAT:

Tyrosine aminotransferase

NP:

Non-pregnant

T1:

First trimester

T2:

Second trimester

T3:

Third trimester

CYP:

Cytochrome P450

DPBS:

Dulbecco’s phosphate-buffered saline

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Funding

This work was supported by National Institute on Drug Abuse grant P01DA032507.

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Authors and Affiliations

  1. Department of Pharmaceutics, University of Washington, P.O. Box 357610, Seattle, WA, 98195, USA

    Madhav Sachar, Edward J. Kelly & Jashvant D. Unadkat

Authors
  1. Madhav Sachar
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  2. Edward J. Kelly
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  3. Jashvant D. Unadkat
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Contributions

Participated in research design: MS, EJK, and JDU. Conducted experiments: MS. Performed data analysis: MS. Wrote or contributed to the manuscript: MS, EJK, and JDU.

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Correspondence to Jashvant D. Unadkat.

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Sachar, M., Kelly, E.J. & Unadkat, J.D. Mechanisms of CYP3A Induction During Pregnancy: Studies in HepaRG Cells. AAPS J 21, 45 (2019). https://doi.org/10.1208/s12248-019-0316-z

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