Molecular Oncology and Immunotherapy, Clinic of General Surgery, 18057 Rostock, Germany.
Institute of Pharmacology and Toxicology, Rostock University Medical Center, 18057 Rostock, Germany.
Int J Mol Sci. 2022 Aug 30;23(17):9861. doi: 10.3390/ijms23179861.
Commonly used intestinal in vitro models are limited in their potential to predict oral drug absorption. They either lack the capability to form a tight cellular monolayer mimicking the intestinal epithelial barrier or the expression of cytochrome P450 3A4 (CYP3A4). The aim of this study was to establish a platform of colorectal cancer patient-derived cell lines for evaluation of human intestinal drug absorption and metabolism. We characterized ten 2D cell lines out of our collection with confluent outgrowth and long-lasting barrier forming potential as well as suitability for high throughput applications with special emphasis on expression and inducibility of CYP3A4. By assessment of the transepithelial electrical resistance (TEER) the cells barrier function capacity can be quantified. Very high TEER levels were detected for HROC60. A high basal CYP3A4 expression and function was found for HROC32. Eight cell lines showed higher CYP3A4 induction by stimulation via the vitamin D receptor compared to Caco-2 cells (5.1- to 16.8-fold change). Stimulation of the pregnane X receptor led to higher CYP3A4 induction in two cell lines. In sum, we identified the two cell lines HROC183 T0 M2 and HROC217 T1 M2 as useful tools for in vitro drug absorption studies. Due to their high TEER values and inducibility by drug receptor ligands, they may be superior to Caco-2 cells to analyze oral drug absorption and intestinal drug-drug interactions. Significance statement: Selecting appropriate candidates is important in preclinical drug development. Therefore, cell models to predict absorption from the human intestine are of the utmost importance. This study revealed that the human cell lines HROC183 T0 M2 and HROC217 T1 M2 may be better suited models and possess higher predictive power of pregnane X receptor- and vitamin D-mediated drug metabolism than Caco-2 cells. Consequently, they represent useful tools for predicting intestinal absorption and simultaneously enable assessment of membrane permeability and first-pass metabolism.
常用的肠道体外模型在预测口服药物吸收方面的潜力有限。它们要么缺乏形成模拟肠道上皮屏障的紧密细胞单层的能力，要么缺乏细胞色素 P450 3A4（CYP3A4）的表达。本研究旨在建立一个结直肠癌患者来源细胞系的平台，用于评估人类肠道药物吸收和代谢。我们从我们的细胞系库中鉴定了 10 种 2D 细胞系，这些细胞系具有融合性生长和持久的屏障形成潜力，并且适合高通量应用，特别强调 CYP3A4 的表达和诱导性。通过评估跨上皮电阻（TEER），可以定量细胞的屏障功能能力。HROC60 的 TEER 水平非常高。HROC32 的 CYP3A4 表达和功能基础较高。与 Caco-2 细胞相比，有 8 种细胞系通过维生素 D 受体刺激显示出更高的 CYP3A4 诱导作用（5.1-16.8 倍变化）。两种细胞系在刺激孕烷 X 受体后，CYP3A4 的诱导作用更高。总的来说，我们鉴定出两种细胞系 HROC183 T0 M2 和 HROC217 T1 M2 是用于体外药物吸收研究的有用工具。由于它们的 TEER 值较高，并且可以被药物受体配体诱导，因此它们可能优于 Caco-2 细胞，可用于分析口服药物吸收和肠道药物相互作用。
在临床前药物开发中，选择合适的候选物非常重要。因此，预测来自人类肠道吸收的细胞模型是最重要的。本研究表明，HROC183 T0 M2 和 HROC217 T1 M2 这两种人类细胞系可能是更好的模型，并且比 Caco-2 细胞具有更高的预测妊娠烷 X 受体和维生素 D 介导的药物代谢能力。因此，它们代表了预测肠道吸收的有用工具，同时能够评估膜通透性和首过代谢。
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