Fashe Muluneh M, Fallon John K, Miner Taryn A, Tiley Jacqueline B, Smith Philip C, Lee Craig R
Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Division of Pharmacoengineering and Molecular Pharmaceutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Front Pharmacol. 2022 Sep 21;13:1004010. doi: 10.3389/fphar.2022.1004010. eCollection 2022.
Pregnancy alters the disposition and exposure to multiple drugs indicated for pregnancy-related complications. Previous studies have shown that pregnancy-related hormones (PRHs) alter the expression and function of certain cytochrome P450s (CYPs) in human hepatocytes. However, the impact of PRHs on hepatic concentrations of non-CYP drug-metabolizing enzymes (DMEs) and transport proteins remain largely unknown. In this study, sandwich-cultured human hepatocytes (SCHH) from five female donors were exposed to vehicle or PRHs (estrone, estradiol, estriol, progesterone, cortisol, and placental growth hormone), administered individually or in combination, across a range of physiologically relevant PRH concentrations for 72 h. Absolute concentrations of 33 hepatic non-CYP DMEs and transport proteins were quantified in SCHH membrane fractions using a quantitative targeted absolute proteomics (QTAP) isotope dilution nanoLC-MS/MS method. The data revealed that PRHs altered the absolute protein concentration of various DMEs and transporters in a concentration-, isoform-, and hepatocyte donor-dependent manner. Overall, eight of 33 (24%) proteins exhibited a significant PRH-evoked net change in absolute protein concentration relative to vehicle control (ANOVA < 0.05) across hepatocyte donors: 1/11 UGTs (9%; UGT1A4), 4/6 other DMEs (67%; CES1, CES2, FMO5, POR), and 3/16 transport proteins (19%; OAT2, OCT3, P-GP). An additional 8 (24%) proteins (UGT1A1, UGT2B4, UGT2B10, FMO3, OCT1, MRP2, MRP3, ENT1) exhibited significant PRH alterations in absolute protein concentration within at least two individual hepatocyte donors. In contrast, 17 (52%) proteins exhibited no discernable impact by PRHs either within or across hepatocyte donors. Collectively, these results provide the first comprehensive quantitative proteomic evaluation of PRH effects on non-CYP DMEs and transport proteins in SCHH and offer mechanistic insight into the altered disposition of drug substrates cleared by these pathways during pregnancy.
妊娠会改变多种用于治疗妊娠相关并发症药物的处置和暴露情况。以往研究表明，妊娠相关激素（PRHs）会改变人肝细胞中某些细胞色素P450（CYPs）的表达和功能。然而，PRHs对非CYP药物代谢酶（DMEs）和转运蛋白肝脏浓度的影响仍 largely未知。在本研究中，来自五名女性供体的三明治培养人肝细胞（SCHH）暴露于载体或PRHs（雌酮、雌二醇、雌三醇、孕酮、皮质醇和胎盘生长激素），单独或联合给药，在一系列生理相关的PRH浓度下处理72小时。使用定量靶向绝对蛋白质组学（QTAP）同位素稀释纳米液相色谱-串联质谱法对SCHH膜组分中33种肝脏非CYP DMEs和转运蛋白的绝对浓度进行定量。数据显示，PRHs以浓度、同工型和肝细胞供体依赖性方式改变了各种DMEs和转运蛋白的绝对蛋白浓度。总体而言，33种蛋白中有8种（24%）在肝细胞供体中相对于载体对照表现出PRH引起的绝对蛋白浓度显著净变化（方差分析<0.05）：1/11种UGTs（9%；UGT1A4）、4/6种其他DMEs（67%；CES1、CES2、FMO5、POR）和3/16种转运蛋白（19%；OAT2、OCT3、P-GP）。另外8种（24%）蛋白（UGT1A1、UGT2B4、UGT2B10、FMO3、OCT1、MRP2、MRP3、ENT1）在至少两个个体肝细胞供体中绝对蛋白浓度表现出显著的PRH改变。相比之下，17种（52%）蛋白在肝细胞供体内或供体间均未表现出PRHs的可察觉影响。总体而言，这些结果首次对PRHs对SCHH中非CYP DMEs和转运蛋白的影响进行了全面的定量蛋白质组学评估，并为孕期这些途径清除的药物底物处置改变提供了机制性见解。
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