Quantitative profiling of the UGT transcriptome in human drug-metabolizing tissues

Tourancheau, Alan; Rouleau, Michèle; Guauque-Olarte, Sandra; Villeneuve, Lyne; Gilbert, Isabelle; Droit, Arnaud; Guillemette, Chantal

doi:10.1038/tpj.2017.5

Cited by 35 publications

(40 citation statements)

References 31 publications

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“…Several previous studies have used RNA expression and enzymatic kinetics as a surrogate to protein quantification (Lazarus et al, 2005;Izukawa et al, 2009;Chen et al, 2010;Jones and Lazarus, 2014;Liu et al, 2014). These methods, however, have inherent limitations, such as the presence of extensive alternative splicing events (Tourancheau et al, 2016(Tourancheau et al, , 2018 and the intrinsic overlapping substrate specificities characterizing these enzymes (Belanger et al, 1998;Guillemette et al, 2010Guillemette et al, , 2014. More recently, to circumvent this technical issue related to the absence of highly specific UGT antibodies, MS-based methods were developed to accurately quantify UGT isoforms in human samples using small signature peptides (Sakamoto et al, 2011;Fallon et al, 2013;Sato et al, 2014;Margaillan et al, 2015b;Bhatt et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Liu et al, 2014). These approaches have limitations, especially in the context of splicing mechanisms and overlapping substrate specificities (Hum et al, 1999;Guillemette et al, 2014;Tourancheau et al, 2016Tourancheau et al, , 2018. More recently, mass spectrometry (MS)-based quantification of UGT2B17 was developed using signature peptides (Fallon et al, 2013;Margaillan et al, 2015b;Bhatt et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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Factors Affecting Interindividual Variability of Hepatic UGT2B17 Protein Expression Examined Using a Novel Specific Monoclonal Antibody

et al. 2019

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Accurate quantification of the metabolic enzyme uridine diphosphoglucuronosyltransferase (UGT) UGT2B17 has been hampered by the high sequence identity with other UGT2B enzymes (as high as 94%) and by the lack of a specific antibody. Knowing the significance of the UGT2B17 pathway in drug and hormone metabolism and cancer, we developed a specific monoclonal antibody (EL-2B17mAb), initially validated by the lack of detection in liver microsomes of an individual carrying no UGT2B17 gene copy and in supersomes expressing UGT2B enzymes. Immunohistochemical detection in livers revealed strong labeling of bile ducts and variable labeling of hepatocytes. Expression levels assessed by immunoblotting were highly correlated to mass spectrometry-based quantification (r = 0.93), and three major expression patterns (absent, low, or high) were evidenced. Livers with very low expression were carriers of the functional rs59678213 G variant, located in the binding site for the transcription factor forkhead box A1 (FOXA1) of the UGT2B17 promoter. The highest level of expression was observed for individuals carrying at least one rs59678213 A allele. Multiple regression analysis indicated that the number of gene copies explained only 8% of UGT2B17 protein expression, 49% when adding rs59678213, reaching 54% when including sex. The novel EL-2B17mAb antibody allowed specific UGT2B17 quantification and exposed different patterns of hepatic expression. It further suggests that FOXA1 is a key driver of UGT2B17 expression in the liver. The availability of this molecular tool will help characterize the UGT2B17 level in various disease states and establish more precisely the contribution of the UGT2B17 enzyme to drug and hormone metabolism.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Factors Affecting Interindividual Variability of Hepatic UGT2B17 Protein Expression Examined Using a Novel Specific Monoclonal Antibody

et al. 2019

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“…The tissue-specific expression of UGTs is highly regulated by multiple signalling pathways and transcription factors, and might be linked to the substrate metabolites they conjugate. 2,3 The substrate molecules can be from endogenous or exogenous sources (Figs. 1c, 2a).…”

Section: Introductionmentioning

confidence: 99%

Emerging roles for UDP-glucuronosyltransferases in drug resistance and cancer progression

et al. 2020

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The best-known role of UDP-glucuronosyltransferase enzymes (UGTs) in cancer is the metabolic inactivation of drug therapies. By conjugating glucuronic acid to lipophilic drugs, UGTs impair the biological activity and enhance the water solubility of these agents, driving their elimination. Multiple clinical observations support an expanding role for UGTs as modulators of the drug response and in mediating drug resistance in numerous cancer types. However, accumulating evidence also suggests an influence of the UGT pathway on cancer progression. Dysregulation of the expression and activity of UGTs has been associated with the progression of several cancers, arguing for UGTs as possible mediators of oncogenic pathways and/or disease accelerators in a drug-naive context. The consequences of altered UGT activity on tumour biology are incompletely understood. They might be associated with perturbed levels of bioactive endogenous metabolites such as steroids and bioactive lipids that are inactivated by UGTs or through non-enzymatic mechanisms, thereby eliciting oncogenic signalling cascades. This review highlights the evidence supporting dual roles for the UGT pathway, affecting cancer progression and drug resistance. Pharmacogenomic testing of UGT profiles in patients and the development of therapeutic options that impair UGT actions could provide useful prognostic and predictive biomarkers and enhance the efficacy of anti-cancer drugs.

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“…In contrast, the involvement of multiple UGT enzymes in the metabolism of IMM-H004 correspondingly makes it less likely that the PK profile of IMM-H004 was affected by other drugs. Since the abundant isoforms of UGT enzyme expression in human liver are UGT2B7, 1A4, 2B4, 1A1, 2B15, and 1A9, followed by UGT1A6 and 1A3, while UGT1A7 and 1A8 are mainly expressed in extrahepatic tissues (Izukawa et al, 2009; Achour et al, 2017; Tourancheau et al, 2018), it is anticipated that UGT1A1 and 1A9 are the major contributors for the formation of IMM-H004G in humans. It has been reported that hydroxycoumarin derivatives were good substrates of UGT.…”

Section: Discussionmentioning

confidence: 99%

Metabolism of IMM-H004 and Its Pharmacokinetic-Pharmacodynamic Analysis in Cerebral Ischemia/Reperfusion Injured Rats

et al. 2019

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IMM-H004, a derivative of coumarin, is a promising candidate for the treatment of cerebral ischemia. The pharmacodynamic mechanisms of IMM-H004 are still under exploration. The present study was conducted to explore the pharmacoactive substances of IMM-H004 from the perspective of drug metabolism. Four metabolites of IMM-H004 including demethylated metabolites M1 and M2, glucuronide conjugate IMM-H004G (M3), and sulfated conjugate M4 were found in rats in vivo . IMM-H004G was the major metabolite in rats and cultured human hepatocytes, and uridine diphosphate-glucuronosyltransferase (UGT) was found to catalyze the metabolism of IMM-H004 in human liver microsomes (HLMs) and rat liver microsomes (RLMs) with high capacity ( V max at 3.25 and 5.04 nmol/min/mg protein). Among 13 recombinant human UGT isoforms, UGT1A7, 1A9, 1A8, and 1A1 appeared to be primarily responsible for IMM-H004G formation. The exposure and duration of IMM-H004G (28,948 h × ng/ml of area under the plasma concentration–time curve (AUC), 6.61 h of t 1/2β ) was much higher than that of the parent drug (1,638 h × ng/ml of AUC, 0.42 h of t 1/2β ) in transient middle cerebral artery occlusion/reperfusion (MCAO/R) rats, consistent with the malondialdehyde (MDA) inhibition effect for at least 10 h. Further pharmacological study revealed that IMM-H004G exhibited a similar neuroprotective activity to that of the parent drug on both oxygen-glucose deprivation injured PC12 cells and transient MCAO/R injured rats. These results demonstrate that both prototype and IMM-H004G are the active pharmaceutical substances, and IMM-H004G, at least in part, contributes to the maintenance of anti-cerebral ischemia efficacy of IMM-H004.

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Quantitative profiling of the UGT transcriptome in human drug-metabolizing tissues

Cited by 35 publications

References 31 publications

Factors Affecting Interindividual Variability of Hepatic UGT2B17 Protein Expression Examined Using a Novel Specific Monoclonal Antibody

Factors Affecting Interindividual Variability of Hepatic UGT2B17 Protein Expression Examined Using a Novel Specific Monoclonal Antibody

Emerging roles for UDP-glucuronosyltransferases in drug resistance and cancer progression

Metabolism of IMM-H004 and Its Pharmacokinetic-Pharmacodynamic Analysis in Cerebral Ischemia/Reperfusion Injured Rats

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