Albadry Mohamed, Küttner Jonas, Grzegorzewski Jan, Dirsch Olaf, Kindler Eva, Klopfleisch Robert, Liska Vaclav, Moulisova Vladimira, Nickel Sandra, Palek Richard, Rosendorf Jachym, Saalfeld Sylvia, Settmacher Utz, Tautenhahn Hans-Michael, König Matthias, Dahmen Uta
Department of General, Visceral and Vascular Surgery, Experimental Transplantation Surgery, Jena University Hospital, Jena, Germany.
Department of Pathology, Faculty of Veterinary Medicine, Menoufia University, Shebin Elkom, Menoufia, Egypt.
Front Pharmacol. 2024 May 16;15:1404938. doi: 10.3389/fphar.2024.1404938. eCollection 2024.
There is a lack of systematic research exploring cross-species variation in liver lobular geometry and zonation patterns of critical drug-metabolizing enzymes, a knowledge gap essential for translational studies. This study investigated the critical interplay between lobular geometry and key cytochrome P450 (CYP) zonation in four species: mouse, rat, pig, and human. We developed an automated pipeline based on whole slide images (WSI) of hematoxylin-eosin-stained liver sections and immunohistochemistry. This pipeline allows accurate quantification of both lobular geometry and zonation patterns of essential CYP proteins. Our analysis of CYP zonal expression shows that all CYP enzymes (besides CYP2D6 with panlobular expression) were observed in the pericentral region in all species, but with distinct differences. Comparison of normalized gradient intensity shows a high similarity between mice and humans, followed by rats. Specifically, CYP1A2 was expressed throughout the pericentral region in mice and humans, whereas it was restricted to a narrow pericentral rim in rats and showed a panlobular pattern in pigs. Similarly, CYP3A4 is present in the pericentral region, but its extent varies considerably in rats and appears panlobular in pigs. CYP2D6 zonal expression consistently shows a panlobular pattern in all species, although the intensity varies. CYP2E1 zonal expression covered the entire pericentral region with extension into the midzone in all four species, suggesting its potential for further cross-species analysis. Analysis of lobular geometry revealed an increase in lobular size with increasing species size, whereas lobular compactness was similar. Based on our results, zonated CYP expression in mice is most similar to humans. Therefore, mice appear to be the most appropriate species for drug metabolism studies unless larger species are required for other purposes, e.g., surgical reasons. CYP selection should be based on species, with CYP2E1 and CYP2D6 being the most preferable to compare four species. CYP1A2 could be considered as an additional CYP for rodent versus human comparisons, and CYP3A4 for mouse/human comparisons. In conclusion, our image analysis pipeline together with suggestions for species and CYP selection can serve to improve future cross-species and translational drug metabolism studies.
目前缺乏系统性研究来探索肝小叶几何结构以及关键药物代谢酶分区模式的跨物种差异，而这一知识空白对于转化研究至关重要。本研究调查了小鼠、大鼠、猪和人类这四个物种中肝小叶几何结构与关键细胞色素P450（CYP）分区之间的关键相互作用。我们基于苏木精-伊红染色肝切片的全玻片图像（WSI）和免疫组织化学开发了一种自动化流程。该流程能够准确量化肝小叶几何结构和必需CYP蛋白的分区模式。我们对CYP分区表达的分析表明，除了具有全小叶表达的CYP2D6外，所有物种的中央周围区域均观察到所有CYP酶，但存在明显差异。归一化梯度强度的比较显示小鼠和人类之间相似度较高，其次是大鼠。具体而言，CYP1A2在小鼠和人类的整个中央周围区域均有表达，而在大鼠中仅限于狭窄的中央周围边缘，在猪中呈全小叶模式。同样，CYP3A4存在于中央周围区域，但其范围在大鼠中差异很大，在猪中呈全小叶模式。CYP2D6分区表达在所有物种中均始终呈现全小叶模式，尽管强度有所不同。CYP2E1分区表达覆盖了所有四个物种的整个中央周围区域并延伸至中区，表明其具有进一步进行跨物种分析的潜力。肝小叶几何结构分析显示，随着物种体型增大，肝小叶尺寸增加，而肝小叶紧密程度相似。基于我们的结果，小鼠中CYP的分区表达与人类最为相似。因此，除非出于其他目的（例如手术原因）需要更大的物种，否则小鼠似乎是药物代谢研究最合适的物种。CYP的选择应基于物种，比较四个物种时，CYP2E1和CYP2D6是最优选的。对于啮齿动物与人类的比较，CYP1A2可被视为额外的CYP，对于小鼠/人类的比较，CYP3A4可被视为额外的CYP。总之，我们的图像分析流程以及关于物种和CYP选择的建议有助于改进未来的跨物种和转化药物代谢研究。
