RRC ID 81271
Author Negoro R, Ouchi A, Deguchi S, Takayama K, Fujita T.
Title Refining Hepatocyte Models to Capture the Impact of CYP2D6*10 Utilizing a PITCh System.
Journal Biol Pharm Bull
Abstract CYP2D6 variants contain various single nucleotide polymorphisms as well as differing levels of metabolic activity. Among these, one of the less active variants CYP2D6*10 (100C > T) is the most prevalent mutation in East Asians, including Japanese. This mutation leads to an amino acid substitution from proline to serine, which reduces the stability of CYP2D6 and consequently decreases its metabolic activity. In this study, we used a genome editing technology called the Precise Integration into Target Chromosome (PITCh) system to stably express six drug-metabolizing enzymes (CYP3A4, POR, uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1), CYP1A2, CYP2C19, CYP2C9, and CYP2D6*10) in HepG2 (CYP2D6*10 KI-HepG2) cells to examine the effect of CYP2D6*10 on drug metabolism prediction. The protein expression levels of CYP2D6 in CYP2D6*10 KI-HepG2 cells were reduced relative to those in the CYP3A4-POR-UGT1A1-CYP1A2-CYP2C19-CYP2C9-CYP2D6 knock-in-HepG2 (CYPs-UGT1A1 KI-HepG2) cells. Consistent with the CYP2D6 protein expression results, CYP2D6 metabolic activity in CYP2D6*10 KI-HepG2 cells was reduced relative to CYPs-UGT1A1 KI-HepG2 cells. We successfully generated CYP2D6*10 KI-HepG2 cells with highly expressed, functional CYP2D6*10, as well as CYP1A2, 2C9, 2C19 and 3A4. CYP2D6*10 KI-HepG2 cells could be an invaluable model for hepatic metabolism and hepatotoxicity studies in East Asians, including Japanese.
Volume 47(8)
Pages 1422-1428
Published 2024-1-1
DOI 10.1248/bpb.b24-00202
PMID 39111864
MeSH Cytochrome P-450 CYP2D6* / genetics Cytochrome P-450 CYP2D6* / metabolism Gene Editing / methods Glucuronosyltransferase / genetics Glucuronosyltransferase / metabolism Hep G2 Cells Hepatocytes* / metabolism Humans Models, Biological Polymorphism, Single Nucleotide
Resource
Human and Animal Cells Hep G2(RCB1648)