Evaluation of mRNA expression of human drug-metabolizing enzymes and transporters in chimeric mouse with humanized liver

Nishimura, Masuhiro; Yoshitsugu, Hiroki; Yokoi, Tsuyoshi; Tateno, Chise; Kataoka, Mikiko; Horie, Tetsuhiro; Yoshizato, Katsutoshi; Naito, Shinsaku

doi:10.1080/00498250500307251

Cited by 54 publications

(36 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many recent publications have unambiguously demonstrated the value of humanized mouse livers for the study of drug metabolism and hepatitis research [7][8][9]21,[23][24][25][26][27][28][29] . However, the albumin-uPA transgenic mouse used for these studies has several major practical limitations.…”

Section: Discussionmentioning

confidence: 99%

Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

et al. 2007

View full text Add to dashboard Cite

Mice that could be highly repopulated with human hepatocytes would have many potential uses in drug development and research applications. The best available model of liver humanization, the uroplasminogen-activator transgenic model, has major practical limitations. To provide a broadly useful hepatic xenorepopulation system, we generated severely immunodeficient, fumarylacetoacetate hydrolase (Fah)-deficient mice. After pretreatment with a urokinaseexpressing adenovirus, these animals could be highly engrafted (up to 90%) with human hepatocytes from multiple sources, including liver biopsies. Furthermore, human cells could be serially transplanted from primary donors and repopulate the liver for at least four sequential rounds. The expanded cells displayed typical human drug metabolism. This system provides a robust platform to produce high-quality human hepatocytes for tissue culture. It may also be useful for testing the toxicity of drug metabolites and for evaluating pathogens dependent on human liver cells for replication.The liver is the principal site for the metabolism of xenobiotic compounds, including medical drugs. Because many hepatic enzymes are species specific, it is necessary to evaluate the metabolism of candidate pharmaceuticals using cultured primary human hepatocytes 1,2 . Today, hepatocytes are isolated primarily from cadaveric organs and then shipped to the location where testing will be performed. The condition (viability and state of differentiation) of hepatocytes from cadaveric sources is highly variable, and many cell preparations are of marginal quality. The availability of high-quality human hepatocytes is further hampered by the fact that they cannot be substantially expanded in tissue culture 3,4 . Hepatocytes from readily available mammalian species, such as the mouse, are not suitable for drug testing because they have a different complement of metabolic enzymes and

show abstract

Section: Discussionmentioning

confidence: 99%

Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

et al. 2007

View full text Add to dashboard Cite

show abstract

“…Furthermore, in the murine environment, these human hepatocytes retained expression of human CYPs and transporter proteins (Katoh and Yokoi, 2007;Nishimura et al, 2005;Tateno et al, 2004;Tsuge et al, 2005) and can be expected to retain human liver function. Although the light and electron microscopic features of the humanized hepatocyte areas showed incompletely developed architecture, sinusoid-like CD31-positive vascular channels, hepatic cord-like structures, and bile canaliculi were observed in the well-differentiated human hepatocyte areas.…”

Section: Toxicological Responses To Apapmentioning

confidence: 99%

Human Hepatocytes Can Repopulate Mouse Liver: Histopathology of the Liver in Human Hepatocyte-Transplanted Chimeric Mice and Toxicologic Responses to Acetaminophen

et al. 2008

View full text Add to dashboard Cite

A human hepatocyte-transplanted chimeric mouse has been established by transplantation of human hepatocytes to urokinase-type plasminogen activator transgenic/severe combined immunodeficiency (uPA +/+ /SCID) mice. These chimeric mice have various amounts of human hepatocytes that proliferate extensively and progressively replace mouse hepatocytes. In the chimeric liver, hepatic cords and sinusoid-like structures were observed. The human hepatocytes expressed human albumin, human cytochrome P450 enzymes, and human transporter proteins. Furthermore, electron microscopic analysis demonstrated bile canaliculi associated with human hepatocytes in the chimeric mouse livers. These results indicate that the chimeric mouse livers contain functionally intact and differentiated human hepatocytes. Additionally, the toxicologic response of hepatocytes to acetaminophen (APAP) administration was compared in normal and chimeric mouse livers. Following 1,400 mg/kg APAP, mild hepatocellular degeneration was observed in the human hepatocyte areas in the chimeric mice, compared with severe centrilobular hepatocellular necrosis in the ICR mouse livers. In conclusion, these chimeric livers contain functionally differentiated human hepatocytes, and are less susceptible to APAP toxicity, compared to ICR mice.

show abstract

“…We considered the possibility that PXB mice, in which hepatocytes are replaced with human hepatocytes to the extent of approximately 80% , may have superior predictive utility, because the expression levels and activities of both P450 and non-P450 enzymes well reflect those of the donor hepatocytes (Yoshitsugu et al, 2006;Yamasaki et al, 2010). In this study, we checked metabolic activities (CYP2C9, CYP2D6, UGT, SULT, and AO) using probe substrates between donor hepatocytes and h-hepatocytes purified from PXB mice (Supplemental Table 3) as well as the expression of drug transporters and blood albumin Nishimura et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Prediction of In Vivo Hepatic Clearance and Half-Life of Drug Candidates in Human Using Chimeric Mice with Humanized Liver

Sanoh

Horiguchi²,

Sugihara³

et al. 2011

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Accurate prediction of pharmacokinetics (PK) parameters in humans from animal data is difficult for various reasons, including species differences. However, chimeric mice with humanized liver (PXB mice; urokinase-type plasminogen activator/severe combined immunodeficiency mice repopulated with approximately 80% human hepatocytes) have been developed. The expression levels and metabolic activities of cytochrome P450 (P450) and non-P450 enzymes in the livers of PXB mice are similar to those in humans. In this study, we examined the predictability for human PK parameters from data obtained in PXB mice. Elimination of selected drugs involves multiple metabolic pathways mediated not only by P450 but also by non-P450 enzymes, such as UDP-glucuronosyltransferase, sulfotransferase, and aldehyde oxidase in liver. ). Elimination half-life (t 1/2 ) after intravenous administration also showed a good correlation (r 2 ‫؍‬ 0.886, p ‫؍‬ 1.506 ؋ 10 ؊4) between humans and PXB mice. The rank order of CL and t 1/2 in human could be predicted at least, although it may not be possible to predict absolute values due to rather large prediction errors. Our results indicate that in vitro and in vivo experiments with PXB mice should be useful at least for semiquantitative prediction of the PK characteristics of candidate drugs in humans.

show abstract

Evaluation of mRNA expression of human drug-metabolizing enzymes and transporters in chimeric mouse with humanized liver

Cited by 54 publications

References 35 publications

Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

Human Hepatocytes Can Repopulate Mouse Liver: Histopathology of the Liver in Human Hepatocyte-Transplanted Chimeric Mice and Toxicologic Responses to Acetaminophen

Prediction of In Vivo Hepatic Clearance and Half-Life of Drug Candidates in Human Using Chimeric Mice with Humanized Liver

Contact Info

Product

Resources

About