Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity

doi:10.1124/dmd.120.000275

	Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity
	Qiao, Shida 1,2; Feng, Sisi 3; Wu, Zhitao 1,2,4; He, Ting 5; Ma, Chen 1,2; Peng, Zhaoliang 1,2; Tian, E.3; Pan, Guoyu 1,2
刊名	DRUG METABOLISM AND DISPOSITION
	2021-04-01
卷号	49 期号:4 页码:305-313
ISSN号	0090-9556
DOI	10.1124/dmd.120.000275
通讯作者	Pan, Guoyu(gypan@simm.ac.cn)
英文摘要	To develop a functional alternative hepatocyte model for primary human hepatocytes (PHHs) with proliferative property, essential drug metabolic, and transporter functions, proliferating human hepatocytes (ProliHHs) expanded from PHHs were fully characterized in vitro. Herein, ProliHHs generated from multiple PHHs donors could be expanded more than 200-fold within four passages and maintained their metabolic or transporter capacities partially. Furthermore, ProliHHs were able to regain the mature hepatic property after three-dimensional (3D) culture. Particularly, the downregulated mRNA expression and function of three major cytochrome P450 (P450) enzymes (CYP1A2, CYP2B6, and CYP3A4) in the proliferating process (ProliHHs-P) could be recovered by 3D culture. The metabolic variabilities across different PHHs donors could be inherited to their matured ProliHHs (ProliHHs-M). The intrinsic clearances of seven major P450 enzymes in ProliHHs-M correlated well (r = 0.87) with those in PHHs. Also, bile canaliculi structures could be observed in sandwich-cultured ProliHHs (SC-ProliHHs), and the biliary excretion index of four probe compounds [cholyl-lysfluorescein, 5-(and-6)-carboxy-29, 79-dichlorofluorescein diacetate (CDF), deuterium-labeled sodiumtaurocholate acid, and rosuvastatin] in SC-ProliHHs (>10%) were close to sandwich-cultured PHHs. More importantly, both ProliHHs-P and ProliHHs-M could be used to evaluate hepatotoxicity. Therefore, these findings demonstrated that the 3D and sandwich culture system could be used to recover the metabolic and transporter functions in ProliHHs for clearance prediction and cholestasis risk assessment, respectively. Together, ProliHHs could be a promising substitute for PHHs in drug metabolism, transport, and hepatotoxicity screening. SIGNIFICANCE STATEMENT This report describes the study of drug metabolic capacities, efflux transporter functions, and toxicity assessments of proliferating human hepatocytes (ProliHHs). The metabolic variability in different primary human hepatocyte donors could be inherited by their matured ProliHHs derivatives. Also, ProliHHs could form canalicular networks in sandwich culture and display biliary excretion capacities. More importantly, both the proliferative and maturation statuses of ProliHHs could be used to evaluate hepatotoxicity. Together, ProliHHs were feasible to support drug candidate screening in hepatic metabolism, disposition, and toxicity.
资助项目	Organ Reconstruction and Manufacturing Strategic Priority Research Program of the Chinese Academy of Sciences[XDA16020205] ; National Science Foundation of China[81872927] ; International Partnership Program of Chinese Academy of Sciences[153631KYSB20160004] ; Independent Deployment Program of the Institute of Pharmaceutical Innovation of the Chinese Academy of Sciences[CASIMM0120184005] ; China Postdoctoral Science Foundation[2019M651623]
WOS研究方向	Pharmacology & Pharmacy
语种	英语
出版者	AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS
WOS记录号	WOS:000637605000003
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/295203]
专题	中国科学院上海药物研究所
通讯作者	Pan, Guoyu
作者单位	1.Chinese Acad Sci, Shanghai Inst Mat Med, Haike Rd 501, Shanghai 201203, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Shanghai Hexaell Biotech Co Ltd, Shanghai, Peoples R China 4.Nanjing Univ Chinese Med, Nanjing, Peoples R China 5.Nanjing Tech Univ, Nanjing, Peoples R China
推荐引用方式 GB/T 7714	Qiao, Shida,Feng, Sisi,Wu, Zhitao,et al. Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity[J]. DRUG METABOLISM AND DISPOSITION,2021,49(4):305-313.
APA	Qiao, Shida.,Feng, Sisi.,Wu, Zhitao.,He, Ting.,Ma, Chen.,...&Pan, Guoyu.(2021).Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity.DRUG METABOLISM AND DISPOSITION,49(4),305-313.
MLA	Qiao, Shida,et al."Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity".DRUG METABOLISM AND DISPOSITION 49.4(2021):305-313.