Human quad liver-on-chip system as a tool toward bridging the gap between animals and humans regarding toxicology and pharmacology of a cannabidiol-rich cannabis extract

Cannabidiol (CBD) is a major phytocannabinoid from <i>Cannabis sativa</i>. It is currently widely available and widely used in the USA, but despite its rapid progress to market, the pharmacology and toxicology of both CBD and cannabidiol-rich cannabis extracts (CRCE) remain largely unknown. The goals of this study were to investigate the potential of a novel human microphysiological system to emulate CRCE-induced hepatotoxicity and pharmacological properties demonstrated in animal models. For this purpose, C57BL6/J male mice were subjected to dosing with either 0, 61.5, 184.5, or 615 mg/kg of CRCE for 10 days. The liver-on-chip system, incorporating human primary hepatocytes, sinusoidal endothelial cells, as well as Kupffer and stellate cells was subjected to 0, 300, 1,200, or 4,400 ng/mL of CRCE (8 h exposure followed by 16 h washout) for 5 days. Administration of CRCE in mice resulted in nearly 4-fold elevations of plasma ALT at 615 mg/kg (<i>p</i> &lt; 0.01) and a dose-dependent decrease in intrahepatic miR-122. Elevated levels of ALT, paralleled by decreased intrahepatic and increased effluent levels of miR-122, were also observed in the liver-on-chip, although these results were not statistically significant. Exposure to CRCE resulted in a robust and dose-dependent induction of key cytochrome P450 enzymes, namely <i>Cyp1a2</i>, <i>Cyp2b6</i> (<i>CYP2B10</i>), <i>Cyp2e1</i>, and <i>Cyp2c9</i> (<i>CYP2C19</i>) in both mouse livers and liver-on-chip. The results of this study demonstrate the congruence between the responses observed in mouse and human liver-on-chip experimental systems and provide evidence of the potential microphysiological systems hold for translating animal data into clinical practice.

大麻二酚（Cannabidiol, CBD）是大麻（Cannabis sativa）中的主要植物大麻素。目前其在美国广泛可及并被广泛使用，尽管其市场化进程迅速，但CBD及富含大麻二酚的大麻提取物（CRCE）的药理学与毒理学特性仍未得到充分阐释。本研究旨在探究一款新型人类微生理系统（human microphysiological system）模拟CRCE诱导的肝毒性，以及复现动物模型中观察到的药理特性的潜力。为此，研究人员对C57BL/6J雄性小鼠分别给予0、61.5、184.5或615 mg/kg的CRCE灌胃给药，持续10天。本研究所用的肝脏芯片器官系统，整合了人原代肝细胞、肝窦内皮细胞、库普弗细胞与肝星状细胞，分别以0、300、1200或4400 ng/mL的CRCE进行处理：先暴露8小时，随后洗脱16小时，持续干预5天。小鼠给药实验结果显示，当给药剂量为615 mg/kg时，血浆丙氨酸氨基转移酶（ALT）水平升高近4倍（P<0.01），且肝内miR-122水平呈剂量依赖性降低。在肝脏芯片器官系统中，同样观察到ALT水平升高，伴随肝内miR-122水平降低与流出液中miR-122水平升高，但上述结果未达到统计学显著性。CRCE暴露可在小鼠肝脏与肝脏芯片器官系统中，强效且剂量依赖性地诱导关键细胞色素P450酶的表达，具体包括Cyp1a2、Cyp2b6（CYP2B10）、Cyp2e1以及Cyp2c9（CYP2C19）。本研究结果证实，小鼠与人类肝脏芯片器官实验系统的应答具有高度一致性，同时证明微生理系统具备将动物实验数据转化为临床实践的潜在应用价值。