Modeling Human Cytochrome P450 2D6 Metabolism and Drug-Drug Interaction by a Novel Panel of Knockout and Humanized Mouse Lines

Scheer, Nico; Kapelyukh, Yury; McEwan, Jillian; Beuger, Vincent; Stanley, Lesley A.; Rode, Anja; Wolf, C. Roland

doi:10.1124/mol.111.075192

Cited by 53 publications

(54 citation statements)

References 43 publications

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“…Nevertheless, it is not possible to differentiate the relative contributions of individual Cyp2d enzymes to dextrorphan formation. Using Cyp2d knock-out mice, it has been shown that Cyp2d enzymes mediate the overall elimination of dextromethorphan in mice (Scheer et al, 2012), but dextromethorphan metabolism has been kinetically characterized only for Cyp2d22 (K m = 250 mM) (Yu and Haining, 2006). The observed K m values for dextrorphan formation in the mouse liver homogenates (2.5-3.5 mM), were ;40-fold lower than that shown for Cyp2d22, suggesting that multiple enzymes contribute to dextrorphan formation in mouse liver.…”

Section: Discussionmentioning

confidence: 99%

Hepatic Cyp2d and Cyp26a1 mRNAs and Activities Are Increased During Mouse Pregnancy

Topletz

Lee

et al. 2012

Drug Metab Dispos

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There is considerable evidence that drug disposition is altered during human pregnancy and based on probe drug studies, CYP2D6 activity increases during human pregnancy. The aim of this study was to determine whether the changes of CYP2D6 activity observed during human pregnancy could be replicated in the mouse, and explore possible mechanisms of increased CYP2D6 activity during pregnancy. Cyp2d11, Cyp2d22, Cyp2d26 and Cyp2d40 mRNA was increased (P < 0.05) on gestational days (GD) 15 and 19 compared with the non-pregnant controls. There was no change (P > 0.05) in Cyp2d9 and Cyp2d10 mRNA. In agreement with the increased Cyp2d mRNA, Cyp2d-mediated dextrorphan formation from dextromethorphan was increased 2.7-fold (P < 0.05) on GD19 (56.8639.4 pmol/min/mg protein) when compared with the non-pregnant controls (20.8611.2 pmol/min/mg protein). An increase in Cyp26a1 mRNA (10-fold) and retinoic acid receptor (Rar)b mRNA (2.8-fold) was also observed during pregnancy. The increase in Cyp26a1 and Rarb mRNA during pregnancy indicates increased retinoic acid signaling in the liver during pregnancy. A putative retinoic acid response element was identified within the Cyp2d40 promoter and the mRNA of Cyp2d40 correlated (P < 0.05) with Cyp26a1 and Rarb. These results show that Cyp2d mRNA is increased during mouse pregnancy and the mouse may provide a suitable model to investigate the mechanisms underlying the increased clearance of CYP2D6 probes observed during human pregnancy. Our findings also suggest that retinoic acid signaling in the liver is increased during pregnancy, which may have broader implications to energy homeostasis in the liver during pregnancy.

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Section: Discussionmentioning

confidence: 99%

Hepatic Cyp2d and Cyp26a1 mRNAs and Activities Are Increased During Mouse Pregnancy

Topletz

Lee

et al. 2012

Drug Metab Dispos

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“…The generation and characterisation of Cyp2cKO, Cyp2dKO, Cyp3aKO, Cyp2c/2d/3aKO, hCYP2D6*2 and hPXR/hCAR/hCYP3A4/3A7 mice has been described previously (19)(20)(21)(22). For the humanized lines, briefly, the hCYP2D6*2 line contains a targeted insertion of an expression cassette containing 9kb of the CYP2D6 promoter, along with all exons, introns and 5' and 3' untranslated regions, into the murine Cyp2d locus, from which all nine functional murine Cyp2d genes have been deleted (20).…”

Section: Animal Lines and Husbandrymentioning

confidence: 99%

Identification of Novel Pathways of Osimertinib Disposition and Potential Implications for the Outcome of Lung Cancer Therapy

MacLeod

Lin

Huang

et al. 2018

Clinical Cancer Research

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2 Abstract PurposeOsimertinib is a third-generation inhibitor of the epidermal growth factor receptor used in treatment of non-small cell lung cancer. A full understanding of its disposition and capacity for interaction with other medications will facilitate its effective use as a single agent and in combination therapy. Experimental designRecombinant cytochrome P450s and liver microsomal preparations were used to identify novel pathways of osimertinib metabolism in vitro. A panel of knockout and mouse lines humanized for pathways of drug metabolism were used to establish the relevance of these pathways in vivo. ResultsAlthough some osimertinib metabolites were similar in mouse and human liver samples there were several significant differences, in particular a marked species difference in the P450s involved. The murine Cyp2d gene cluster played a predominant role in mouse, whereas CYP3A4 was the major human enzyme responsible for osimertinib metabolism. Induction of this enzyme in CYP3A4 humanized mice substantially decreased circulating osimertinib exposure. Importantly, we discovered a further novel pathway of osimertinib disposition involving CPY1A1. Modulation of CYP1A1/CYP1A2 levels markedly reduced parent drug concentrations, significantly altering metabolite pharmacokinetics (PK) in humanized mice in vivo. ConclusionWe demonstrate that a P450 enzyme expressed in smokers' lungs and lung tumors has the capacity to metabolise osimertinib. This could be a significant factor in defining the outcome of osimertinib treatment. This work also illustrates how P450-humanized mice can be used to identify and mitigate Acquired resistance to first and second-generation EGFR inhibitors usually involves a further EGFR mutation, T790M. Osimertinib (AZD9291) is the most effective treatment in these latter cases and, due to its low toxicity and high level of brain penetration, is currently being explored as a first-line therapy for metastatic disease. Osimertinib is a cytochrome P450 substrate. These enzymes can define both systemic exposure and intra-tumoral drug concentrations and can therefore be major determinants in the outcome of cancer therapy. We report a novel pathway of osimertinib disposition which may be of importance in this regard.Research.on May 9, 2018.

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“…However, the single-Cyp knockout mouse models are often unsatisfactory due to the multiplicity of Cyp genes in many of the mouse Cyp subfamilies and to the overlapping substrate specificity of the corresponding P450 enzymes. Alternative strategies that would delete multiple Cyp genes (e.g., those of the Cyp3a subfamily (van Herwaarden et al, 2007) and Cyp2d subfamily (Scheer et al, 2012), knockdown the expression of multiple Cyp genes (Damiri et al, 2012), or abolish all microsomal P450 activities through deletion or down-regulation of the Cpr gene (e.g., Gu et al, 2003;Wu et al, 2005;Wei et al, 2010) have been reported, and the resultant mouse models have been found to have unique advantages for functional studies (e.g., Gu et al, 2005;Weng et al, 2007;Conroy et al, 2010). In this study, we have generated a novel Cyp2a(4/5)bgs-null strain in which nine of the Cyp genes in a 1.2-megabase pair (Mb) Cyp2 cluster in mouse chromosome 7 (including,sequentially,Cyp2a5,2g1,2b19,2b23,2a4,2b9,2b13,2b10, and 2s1) are removed by using the CreloxP technology (Nagy, 2000).…”

Section: Introductionmentioning

confidence: 99%

Generation and Characterization of a Novel Cyp2a(4/5)bgs-Null Mouse Model

Wei

Zhou

et al. 2012

Drug Metab Dispos

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Knockout mouse models targeting various cytochrome P450 (P450 or CYP) genes are valuable for determining P450's biologic functions, including roles in drug metabolism and chemical toxicity. In this study, a novel Cyp2a(4/5)bgs-null mouse model was generated, in which a 1.2-megabase pair genomic fragment containing nine Cyp genes in mouse chromosome 7 (including, sequentially, Cyp2a5, 2g1, 2b19, 2b23, 2a4, 2b9, 2b13, 2b10, and 2s1) are deleted, through Cre-mediated recombination in vivo. The resultant mouse strain was viable and fertile, without any developmental deficits or morphologic abnormalities. Deletion of the constitutive genes in the cluster was confirmed by polymerase chain reaction analysis of the genes and the mRNAs in tissues known to express each gene. The loss of this gene cluster led to significant decreases in microsomal activities toward testosterone hydroxylation in various tissues examined, including olfactory mucosa (OM), lung, liver, and brain. In addition, systemic clearance of pentobarbital was decreased in Cyp2a(4/5)bgs-null mice, as indicated by >60% increases in pentobarbital-induced sleeping time, compared with wild-type (WT) mice. This novel Cyp2a(4/5) bgs-null mouse model will be valuable for in vivo studies of drug metabolism and chemical toxicities in various tissues, including the liver, lung, brain, intestine, kidney, skin, and OM, where one or more of the targeted Cyp genes are known to be expressed in WT mice. The model will also be valuable for preparation of humanized mice that express human CYP2A6, CYP2A13, CYP2B6, or CYP2S1, and as a knockout mouse model for five non-P450 genes (Vmn1r184, Nalp9c, Nalp4a, Nalp9a, and Vmn1r185) that were also deleted.

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Modeling Human Cytochrome P450 2D6 Metabolism and Drug-Drug Interaction by a Novel Panel of Knockout and Humanized Mouse Lines

Cited by 53 publications

References 43 publications

Hepatic Cyp2d and Cyp26a1 mRNAs and Activities Are Increased During Mouse Pregnancy

Hepatic Cyp2d and Cyp26a1 mRNAs and Activities Are Increased During Mouse Pregnancy

Identification of Novel Pathways of Osimertinib Disposition and Potential Implications for the Outcome of Lung Cancer Therapy

Generation and Characterization of a Novel Cyp2a(4/5)bgs-Null Mouse Model

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