Department of Development and Regeneration, Stem Cell Institute, KU Leuven, Leuven, Belgium.
Laboratory of Cellular Metabolism and Metabolic Regulation, VIB Center for Cancer Biology, VIB, Leuven, Belgium.
Nat Commun. 2020 Mar 13;11(1):1393. doi: 10.1038/s41467-020-15058-6.
Predicting drug-induced liver injury in a preclinical setting remains challenging, as cultured primary human hepatocytes (PHHs), pluripotent stem cell-derived hepatocyte-like cells (HLCs), and hepatoma cells exhibit poor drug biotransformation capacity. We here demonstrate that hepatic functionality depends more on cellular metabolism and extracellular nutrients than on developmental regulators. Specifically, we demonstrate that increasing extracellular amino acids beyond the nutritional need of HLCs and HepG2 cells induces glucose independence, mitochondrial function, and the acquisition of a transcriptional profile that is closer to PHHs. Moreover, we show that these high levels of amino acids are sufficient to drive HLC and HepG2 drug biotransformation and liver-toxin sensitivity to levels similar to those in PHHs. In conclusion, we provide data indicating that extracellular nutrient levels represent a major determinant of cellular maturity and can be utilized to guide stem cell differentiation to the hepatic lineage.
在临床前环境中预测药物性肝损伤仍然具有挑战性，因为培养的原代人肝细胞（PHH）、多能干细胞衍生的肝细胞样细胞（HLC）和肝癌细胞表现出较差的药物生物转化能力。我们在此证明，肝功能更多地取决于细胞代谢和细胞外营养物质，而不是发育调节剂。具体而言，我们证明了将细胞外氨基酸增加到超过 HLC 和 HepG2 细胞的营养需求以上，会诱导葡萄糖独立性、线粒体功能，并获得更接近 PHH 的转录谱。此外，我们还表明，这些高浓度的氨基酸足以驱动 HLC 和 HepG2 的药物生物转化和肝毒素敏感性，达到与 PHH 相似的水平。总之，我们提供的数据表明，细胞外营养水平是细胞成熟的主要决定因素，可以用于指导干细胞向肝谱系分化。
