TY - JOUR
T1 - Trans-chromosomic mice containing a human CYP3A cluster for prediction of xenobiotic metabolism in humans
AU - Kazuki, Yasuhiro
AU - Kobayashi, Kaoru
AU - Aueviriyavit, Sasitorn
AU - Oshima, Takeshi
AU - Kuroiwa, Yoshimi
AU - Tsukazaki, Yasuko
AU - Senda, Naoto
AU - Kawakami, Hiroki
AU - Ohtsuki, Sumio
AU - Abe, Satoshi
AU - Takiguchi, Masato
AU - Hoshiya, Hidetoshi
AU - Kajitani, Naoyo
AU - Takehara, Shoko
AU - Kubo, Kinya
AU - Terasaki, Tetsuya
AU - Chiba, Kan
AU - Tomizuka, Kazuma
AU - Oshimura, Mitsuo
N1 - Funding Information:
This study was supported in part by the City Area Program (Basic Stage) and the Regional Innovation Strategy Support Program from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) (M.O.), Takeda Science Foundation (M.O.), the 21st Century Center of Excellence (COE) program from Japan Society for the Promotion of Science (JSPS) (M.O.) and Funding Program for Next Generation World-Leading Researchers (NEXT Program) from Japan Society for the Promotion of Science (JSPS) (Y.K.).
PY - 2013/2
Y1 - 2013/2
N2 - Human CYP3A is the most abundant P450 isozyme present in the human liver and small intestine, and metabolizes around 50% of medical drugs on the market. The human CYP3A subfamily comprises four members (CYP3A4, CYP3A5, CYP3A7, CYP3A43) encoded on human chromosome 7. However, transgenic mouse lines carrying the entire human CYP3A cluster have not been constructed because of limitations in conventional cloning techniques. Here, we show that the introduction of a human artificial chromosome (HAC) containing the entire genomic human CYP3A locus recapitulates tissue- and stage-specific expression of human CYP3A genes and xenobiotic metabolism in mice. About 700 kb of the entire CYP3A genomic segment was cloned into a HAC (CYP3A-HAC), and trans-chromosomic (Tc) mice carrying a single copy of germline-transmittable CYP3A-HAC were generated via a chromosome-engineering technique. The tissue- and stage-specific expression profiles of CYP3A genes were consistent with those seen in humans. We further generated mice carrying the CYP3A-HAC in the background homozygous for targeted deletion of most endogenous Cyp3a genes. In this mouse strain with 'fully humanized' CYP3A genes, the kinetics of triazolam metabolism, CYP3A-mediated mechanism-based inactivation effects and formation of fetal-specific metabolites of dehydroepiandrosterone observed in humans were well reproduced. Thus, these mice are likely to be valuable in evaluating novel drugs metabolized by CYP3A enzymes and in studying the regulation of human CYP3A gene expression. Furthermore, this system can also be used for generating Tc mice carrying other human metabolic genes.
AB - Human CYP3A is the most abundant P450 isozyme present in the human liver and small intestine, and metabolizes around 50% of medical drugs on the market. The human CYP3A subfamily comprises four members (CYP3A4, CYP3A5, CYP3A7, CYP3A43) encoded on human chromosome 7. However, transgenic mouse lines carrying the entire human CYP3A cluster have not been constructed because of limitations in conventional cloning techniques. Here, we show that the introduction of a human artificial chromosome (HAC) containing the entire genomic human CYP3A locus recapitulates tissue- and stage-specific expression of human CYP3A genes and xenobiotic metabolism in mice. About 700 kb of the entire CYP3A genomic segment was cloned into a HAC (CYP3A-HAC), and trans-chromosomic (Tc) mice carrying a single copy of germline-transmittable CYP3A-HAC were generated via a chromosome-engineering technique. The tissue- and stage-specific expression profiles of CYP3A genes were consistent with those seen in humans. We further generated mice carrying the CYP3A-HAC in the background homozygous for targeted deletion of most endogenous Cyp3a genes. In this mouse strain with 'fully humanized' CYP3A genes, the kinetics of triazolam metabolism, CYP3A-mediated mechanism-based inactivation effects and formation of fetal-specific metabolites of dehydroepiandrosterone observed in humans were well reproduced. Thus, these mice are likely to be valuable in evaluating novel drugs metabolized by CYP3A enzymes and in studying the regulation of human CYP3A gene expression. Furthermore, this system can also be used for generating Tc mice carrying other human metabolic genes.
UR - http://www.scopus.com/inward/record.url?scp=84872385005&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84872385005&partnerID=8YFLogxK
U2 - 10.1093/hmg/dds468
DO - 10.1093/hmg/dds468
M3 - Article
C2 - 23125282
AN - SCOPUS:84872385005
VL - 22
SP - 578
EP - 592
JO - Human Molecular Genetics
JF - Human Molecular Genetics
SN - 0964-6906
IS - 3
M1 - dds468
ER -