Uehara Shotaro, Higuchi Yuichiro, Yoneda Nao, Kawai Kenji, Yamamoto Masafumi, Kamimura Hidetaka, Iida Yuichi, Oshimura Mitsuo, Kazuki Yasuhiro, Yamazaki Hiroshi, Hikita Hayato, Takehara Tetsuo, Suemizu Hiroshi
Laboratory Animal Research Department, Japan.
Pathology Analysis Center, Japan.
Drug Metab Pharmacokinet. 2022 Feb;42:100410. doi: 10.1016/j.dmpk.2021.100410. Epub 2021 Jun 12.
We developed a novel immunodeficient NOG mouse expressing HSVtk mutant clone 30 cDNA under the control of mouse transthyretin gene enhancer/promoter (NOG-TKm30) to acquire fertility in males and high inducibility of liver injury in females. Maximum human albumin levels (approx. 15 mg/mL plasma) in both male and female NOG-TKm30 mice engrafted with human hepatocytes (humanized liver mice) were observed 8-12 weeks after transplantation. Immunohistochemical analyses revealed abundant expression of major human cytochrome P450 (CYP) enzymes (CYP1A2, CYP2C9, CYP2D6, CYP2E1, and CYP3A4) in reconstituted liver with original zonal distribution. In vivo drug-drug interactions were observed in humanized liver mice as decreased area under the curve of midazolam (CYP3A4/5 substrate) and omeprazole (CYP3A4/5 and CYP2C19 substrate) after oral administration of rifampicin. Furthermore, we developed a pregnant model for evaluating prenatal exposure to drugs. The detection of thalidomide metabolites in the fetuses of pregnant humanized liver mice indicates that the novel TK model can be used for developmental toxicity studies requiring the assessment of human drug metabolism. These results suggest that the limitations of traditional TK-NOG mice can be addressed using NOG-TKm30 mice, which constitute a novel platform for humanized liver for both in vivo and in vitro studies.
我们开发了一种新型免疫缺陷型NOG小鼠，其在小鼠甲状腺转运蛋白基因增强子/启动子的控制下表达单纯疱疹病毒胸苷激酶（HSVtk）突变体克隆30 cDNA（NOG-TKm30），以获得雄性生育能力和雌性肝脏损伤的高诱导性。在移植人肝细胞的雄性和雌性NOG-TKm30小鼠（人源化肝脏小鼠）中，移植后8-12周观察到最大人白蛋白水平（约15 mg/mL血浆）。免疫组织化学分析显示，主要的人细胞色素P450（CYP）酶（CYP1A2、CYP2C9、CYP2D6、CYP2E1和CYP3A4）在重建肝脏中呈原始区域分布且表达丰富。在人源化肝脏小鼠中观察到体内药物相互作用，口服利福平后，咪达唑仑（CYP3A4/5底物）和奥美拉唑（CYP3A4/5和CYP2C19底物）的曲线下面积减小。此外，我们开发了一种用于评估产前药物暴露的妊娠模型。在妊娠人源化肝脏小鼠胎儿中检测到沙利度胺代谢物，表明该新型TK模型可用于需要评估人药物代谢的发育毒性研究。这些结果表明，使用NOG-TKm30小鼠可以解决传统TK-NOG小鼠的局限性，NOG-TKm30小鼠构成了一个用于体内和体外研究的新型人源化肝脏平台。
