High-Throughput Production of Diverse Xenobiotic Metabolites with Cytochrome P450-Transduced Huh7 Hepatoma Cell Lines.

Document Type

Article

Publication Date

9-1-2022

Publication Title

Drug metabolism and disposition: the biological fate of chemicals

Keywords

JGM, Bupropion, Carcinoma, Hepatocellular, Cell Line, Cytochrome P-450 Enzyme System, Humans, Liver Neoplasms, Xenobiotics

JAX Source

Drug Metab Dispos. 2022;50(9):1182-9.

Volume

50

Issue

9

First Page

1182

Last Page

1189

ISSN

1521-009X

PMID

35752443

DOI

https://doi.org/10.1124/dmd.122.000900

Grant

This work was supported by the National Institutes of Health of Environmental Health Science [Grant U2C-ES030163] (to G.W.M., S.L., D.P.J., E.T.M.) and [Grant 1P30-ES019776] (D.P.J., E.T.M).

Abstract

Precision medicine and exposomics require methods to assess xenobiotic metabolism in human metabolomic analyses, including the identification of known and undocumented drug and chemical exposures as well as their metabolites. Recent work demonstrated the use of high-throughput generation of xenobiotic metabolites with human liver S-9 fractions for their detection in human plasma and urine. Here, we tested whether a panel of lentivirally transduced human hepatoma cell lines (Huh7) that express individual cytochrome P450 (P450) enzymes could be used to generate P450-specific metabolites in a high-throughput manner, while simultaneously identifying the enzymes responsible. Cell-line activities were verified using P450-specific probe substrates. To increase analytical throughput, we used a pooling strategy where 36 chemicals were grouped into 12 unique mixtures, each mixture containing 6 randomly selected compounds, and each compound being present in two separate mixtures. Each mixture was incubated with 8 different P450 cell lines for 0 and 2 hours and extracts were analyzed using liquid chromatography-high-resolution mass spectrometry. Cell lines selectively metabolized test substrates, e.g., pazopanib, bupropion, and

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