Absolute Membrane Protein Abundance of P-gp, BCRP and MRPs in Term Human Placenta Tissue and Commonly Used Cell Systems: Application in PBPK Modeling of Placental Drug Disposition.

IF 4.4 3区 医学 Q1 PHARMACOLOGY & PHARMACY
Zubida M Al-Majdoub, Jolien J M Freriksen, Angela Colbers, Jeroen van den Heuvel, Jan Koenderink, Khaled Abduljalil, Brahim Achour, Jill Barber, Rick Greupink, Amin Rostami-Hodjegan
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引用次数: 0

Abstract

The placenta acts as a barrier, excluding noxious substances whilst actively transferring nutrients to the fetus, mediated by various transporters. This study quantified the expression of key placental transporters in term human placenta (n=5) and BeWo, BeWo b30, and JEG-3 placenta cell lines. Combining these results with pregnancy physiologically-based pharmacokinetic (PBPK) modeling, we demonstrate the utility of proteomic analysis for predicting placental drug disposition and fetal exposure. Using targeted proteomics with QconCAT standards, we found significant expression of P-gp, BCRP, MRP2, MRP4, and MRP6 in the human placenta (0.05 - 0.25 pmol/mg membrane protein) with only regional differences observed for P-gp. Unexpectedly, both P-gp and BCRP were below the limit of quantification in the regularly used BeWo cells, indicating that this cell line may not be suitable for the study of placental P-gp and BCRP-mediated transport. In cellular and vesicular overexpression systems, P-gp and BCRP were detectable as expected. Vesicle batches showed consistent P-gp expression correlating with functional activity (N-methyl-quinidine (NMQ) transport). However, BCRP activity (Estrone 3-sulfate (E1S) transport) did not consistently align with expression levels. Incorporating in vitro transporter kinetic data, along with placental transporter abundance, into a PBPK model enabled the evaluation of fetal exposure. Simulation with a hypothetical drug indicated that estimating fetal exposure relies on the intrinsic clearances of relevant transporters. To minimize interlaboratory discrepancies, expression data was generated using consistent proteomic methodologies in the same lab. Integration of this data in pregnancy-PBPK modeling offers a promising tool to investigate maternal, placental and fetal drug exposure. Significance Statement This study quantified the expression of key transporters in human placenta and various placental cell lines, revealing significant expression variations. By integrating these data with PBPK modeling, the study highlights the importance of transporter abundance data in understanding and predicting placental drug disposition.

人胎盘组织和常用细胞系统中 P-gp、BCRP 和 MRPs 的绝对膜蛋白丰度:胎盘药物处置 PBPK 模型中的应用。
胎盘起着屏障的作用,在排除有害物质的同时,通过各种转运体积极地向胎儿输送营养物质。本研究定量检测了足月人类胎盘(n=5)以及 BeWo、BeWo b30 和 JEG-3 胎盘细胞系中主要胎盘转运体的表达。将这些结果与妊娠生理药代动力学(PBPK)模型相结合,我们证明了蛋白质组学分析在预测胎盘药物处置和胎儿暴露方面的实用性。通过使用 QconCAT 标准的靶向蛋白质组学,我们发现 P-gp、BCRP、MRP2、MRP4 和 MRP6 在人类胎盘中有显著表达(0.05 - 0.25 pmol/mg 膜蛋白),只有 P-gp 存在区域差异。意外的是,在常用的 BeWo 细胞中,P-gp 和 BCRP 均低于定量限,这表明该细胞系可能不适合研究胎盘 P-gp 和 BCRP 介导的转运。在细胞和囊泡过表达系统中,可以检测到预期的 P-gp 和 BCRP。囊泡批次显示出与功能活性(N-甲基喹啶(NMQ)转运)相关的一致的 P-gp 表达。然而,BCRP 的活性(3-硫酸雌酮(E1S)转运)与表达水平并不一致。将体外转运体动力学数据和胎盘转运体丰度纳入 PBPK 模型,可以评估胎儿的暴露情况。假定药物的模拟表明,胎儿暴露量的估算依赖于相关转运体的内在清除率。为了尽量减少实验室间的差异,在同一实验室使用一致的蛋白质组学方法生成了表达数据。将这些数据整合到妊娠-PBPK 模型中,为研究母体、胎盘和胎儿的药物暴露提供了一种很有前景的工具。意义声明 本研究量化了人类胎盘和各种胎盘细胞系中关键转运体的表达,揭示了显著的表达变化。通过将这些数据与 PBPK 模型相结合,该研究强调了转运体丰度数据在理解和预测胎盘药物处置方面的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
6.50
自引率
12.80%
发文量
128
审稿时长
3 months
期刊介绍: An important reference for all pharmacology and toxicology departments, DMD is also a valuable resource for medicinal chemists involved in drug design and biochemists with an interest in drug metabolism, expression of drug metabolizing enzymes, and regulation of drug metabolizing enzyme gene expression. Articles provide experimental results from in vitro and in vivo systems that bring you significant and original information on metabolism and disposition of endogenous and exogenous compounds, including pharmacologic agents and environmental chemicals.
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