Department of Pharmacology and Therapeutics, MRC Centre of Drug Safety Science, University of Liverpool, The Sherrington Building, Ashton Street, Liverpool, L69 3GE, UK.
Arch Toxicol. 2021 Jul;95(7):2413-2430. doi: 10.1007/s00204-021-03075-3. Epub 2021 May 29.
Hepatic organoids are a recent innovation in in vitro modeling. Initial studies suggest that organoids better recapitulate the liver phenotype in vitro compared to pre-existing proliferative cell models. However, their potential for drug metabolism and detoxification remains poorly characterized, and their global proteome has yet to be compared to their tissue of origin. This analysis is urgently needed to determine what gain-of-function this new model may represent for modeling the physiological and toxicological response of the liver to xenobiotics. Global proteomic profiling of undifferentiated and differentiated hepatic murine organoids and donor-matched livers was, therefore, performed to assess both their similarity to liver tissue, and the expression of drug-metabolizing enzymes and transporters. This analysis quantified 4405 proteins across all sample types. Data are available via ProteomeXchange (PXD017986). Differentiation of organoids significantly increased the expression of multiple cytochrome P450, phase II enzymes, liver biomarkers and hepatic transporters. While the final phenotype of differentiated organoids is distinct from liver tissue, the organoids contain multiple drug metabolizing and transporter proteins necessary for liver function and drug metabolism, such as cytochrome P450 3A, glutathione-S-transferase alpha and multidrug resistance protein 1A. Indeed, the differentiated organoids were shown to exhibit increased sensitivity to midazolam (10-1000 µM) and irinotecan (1-100 µM), when compared to the undifferentiated organoids. The predicted reduced activity of HNF4A and a resulting dysregulation of RNA polymerase II may explain the partial differentiation of the organoids. Although further experimentation, optimization and characterization is needed relative to pre-existing models to fully contextualize their use as an in vitro model of drug-induced liver injury, hepatic organoids represent an attractive novel model of the response of the liver to xenobiotics. The current study also highlights the utility of global proteomic analyses for rapid and accurate evaluation of organoid-based test systems.
肝类器官是体外建模的一项新创新。初步研究表明，与现有的增殖细胞模型相比，类器官在体外更好地再现了肝脏表型。然而，它们在药物代谢和解毒方面的潜力尚未得到充分描述，其整体蛋白质组尚未与它们的组织来源进行比较。为了确定这种新模型在模拟肝脏对外来物质的生理和毒理反应方面可能具有哪些功能增益，迫切需要进行这项分析。因此，对未分化和分化的肝鼠类器官以及供体匹配的肝脏进行了全局蛋白质组学分析，以评估它们与肝脏组织的相似性，以及药物代谢酶和转运蛋白的表达。该分析量化了所有样本类型的 4405 种蛋白质。数据可通过 ProteomeXchange（PXD017986）获得。类器官的分化显著增加了多种细胞色素 P450、二期酶、肝脏生物标志物和肝脏转运蛋白的表达。虽然分化后的类器官的终表型与肝脏组织不同，但类器官包含多种用于肝脏功能和药物代谢的药物代谢和转运蛋白，如细胞色素 P450 3A、谷胱甘肽 S-转移酶α和多药耐药蛋白 1A。事实上，与未分化的类器官相比，分化的类器官对咪达唑仑（10-1000μM）和伊立替康（1-100μM）表现出更高的敏感性。HNF4A 预测活性降低和 RNA 聚合酶 II 的失调可能解释了类器官的部分分化。尽管相对于现有的模型还需要进一步的实验、优化和特征描述，以充分将其作为药物诱导的肝损伤的体外模型使用，但肝类器官代表了一种有吸引力的肝脏对外来物质反应的新型模型。本研究还强调了全局蛋白质组学分析在快速准确评估基于类器官的测试系统方面的效用。
