Comparative Analysis of Species-Specific Hepatocyte Function and Drug Effects in a Liver Microphysiological System PhysioMimix LC12 and 96-Well Plates

Drug-induced liver
injury (DILI) remains a challenge in drug development,
and interspecies differences in liver toxicity represent a need where
comparative analyses may inform preclinical safety study design. In
vitro testing for species-specific liver effects, especially in complex
models such as microphysiological systems (MPS), may help predict
toxicity before advancing from animal to human studies, or derisk
spurious findings in preclinical species. This study assessed the
utility of the perfusion-based PhysioMimix LC12 MPS as compared to
2D cultures and evaluated species-specific DILI using primary hepatocytes
from human, monkey, rat, and dog. Functional, phenotypic, and transcriptional
profiles were evaluated for up to 14 days. Also, cells were exposed
to species-specific hepatotoxicants such as bosentan (BOS), fialuridine
(FIAU), and a common hepatotoxicant for all species, chlorpromazine
(CPZ)in both PhysioMimix LC12 and traditional 2D cultures.
Hepatocytes in PhysioMimix LC12 showed more stable albumin and urea
production as compared to 2D cultures. Concentration–response
studies with CPZ, BOS, and FIAU were performed in 2D; then, repeated
(5 × every 2 days) exposures to sub-100 × Cmax concentrations were tested in PhysioMimix LC12. Species-specific
differences in cellular and molecular effects of the drugs were observed
in both models; data from PhysioMimix LC12 were reflective of the
expected effects in both animals and humans. Still, variability and
low throughput are limitations of MPS for prospective studies of species-specific
responses. Overall, this study confirms the utility of liver safety
studies using PhysioMimix LC12 and also provides suggestions for experimental
designs to overcome the limitations of more complex test systems.

药物性肝损伤（Drug-induced liver injury, DILI）仍是药物研发过程中的一大挑战，而肝脏毒性的种属差异正是一个亟需通过比较分析为临床前安全性研究设计提供参考的研究方向。针对种属特异性肝脏效应的体外检测，尤其是在微生理系统（Microphysiological Systems, MPS）这类复杂模型中开展的检测，有助于在从动物实验推进至人体研究前预判药物毒性，同时也可规避临床前动物实验中出现假阳性结果的风险。
本研究评估了灌流型PhysioMimix LC12微生理系统相较于二维培养模型的应用价值，并利用人、猴、大鼠、犬的原代肝细胞对种属特异性DILI进行了评价。研究对细胞的功能、表型及转录组特征进行了长达14天的检测分析。此外，研究在PhysioMimix LC12与传统二维培养模型中，分别将细胞暴露于种属特异性肝毒性化合物（如波生坦（Bosentan, BOS）、氟拉夫定（Fialuridine, FIAU））以及所有种属共有的肝毒性化合物氯丙嗪（Chlorpromazine, CPZ）。
相较于二维培养模型，PhysioMimix LC12中的原代肝细胞展现出更为稳定的白蛋白与尿素合成能力。研究先在二维培养模型中开展了氯丙嗪、波生坦及氟拉夫定的浓度-效应关系研究；随后在PhysioMimix LC12模型中，测试了细胞反复（每2天一次，共5次）暴露于低于100倍峰浓度（Cmax）的化合物时的反应。
两种模型中均观测到了药物在细胞与分子层面效应的种属差异；PhysioMimix LC12模型得到的数据能够反映动物与人体中的预期效应。不过，针对种属特异性反应的前瞻性研究中，微生理系统仍存在实验变异大、通量较低的局限。综上，本研究证实了使用PhysioMimix LC12开展肝脏安全性研究的应用价值，并为优化实验设计以克服复杂测试系统的局限性提供了参考建议。