Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Pittsburgh Liver Research Center, Human Synthetic Liver Biology Core, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Int J Mol Sci. 2023 Aug 29;24(17):13406. doi: 10.3390/ijms241713406.
Metabolic-dysfunction-associated steatotic liver disease (MASLD), which affects 30 million people in the US and is anticipated to reach over 100 million by 2030, places a significant financial strain on the healthcare system. There is presently no FDA-approved treatment for MASLD despite its public health significance and financial burden. Understanding the connection between point mutations, liver enzymes, and MASLD is important for comprehending drug toxicity in healthy or diseased individuals. Multiple genetic variations have been linked to MASLD susceptibility through genome-wide association studies (GWAS), either increasing MASLD risk or protecting against it, such as rs738409, rs641738, rs780094, rs72613567, and rs2642438. As the impact of genetic variants on the levels of drug-metabolizing cytochrome P450 (CYP) enzymes in human hepatocytes has not been thoroughly investigated, this study aims to describe the analysis of metabolic functions for selected phase I and phase II liver enzymes in human hepatocytes. For this purpose, fresh isolated primary hepatocytes were obtained from healthy liver donors (n = 126), and liquid chromatography-mass spectrometry (LC-MS) was performed. For the cohorts, participants were classified into minor homozygotes and nonminor homozygotes (major homozygotes + heterozygotes) for five gene polymorphisms. For phase I liver enzymes, we found a significant difference in the activity of CYP1A2 in human hepatocytes carrying ( 0.011) and of CYP2C8 in human hepatocytes carrying ( = 0.004). It was also observed that the activity of CYP2C9 was significantly lower in human hepatocytes carrying ( = 0.001) minor homozygous compared to nonminor homozygous. No significant difference in activity of CYP2E1, CYP2C8, CYP2D6, CYP2E1, CYP3A4, ECOD, FMO, MAO, AO, and CES2 and in any of the phase II liver enzymes between human hepatocytes carrying genetic variants for rs738409, rs641738, GCKR rs780094, rs72613567, and rs2642438 were observed. These findings offer a preliminary assessment of the influence of genetic variations on drug-metabolizing cytochrome P450 (CYP) enzymes in healthy human hepatocytes, which may be useful for future drug discovery investigations.
代谢功能障碍相关脂肪性肝病（MASLD）影响美国 3000 万人，预计到 2030 年将超过 1 亿人，给医疗保健系统带来了巨大的财政压力。尽管 MASLD 具有公共卫生意义和财政负担，但目前还没有 FDA 批准的治疗方法。了解点突变、肝酶与 MASLD 之间的关系对于理解健康个体或患病个体的药物毒性很重要。通过全基因组关联研究（GWAS）发现了多种与 MASLD 易感性相关的遗传变异，这些变异要么增加 MASLD 风险，要么起到保护作用，例如 rs738409、rs641738、rs780094、rs72613567 和 rs2642438。由于遗传变异对人肝细胞中药物代谢细胞色素 P450（CYP）酶水平的影响尚未得到充分研究，因此本研究旨在描述人肝细胞中选定的 I 相和 II 相肝酶的代谢功能分析。为此，从健康供体肝中获得新鲜分离的原代肝细胞（n=126），并进行液相色谱-质谱（LC-MS）分析。在这些队列中，根据五个基因多态性，将参与者分为次要纯合子和非次要纯合子（主要纯合子+杂合子）。对于 I 相肝酶，我们发现携带 rs738409 的人肝细胞中 CYP1A2 的活性存在显著差异（ 0.011），携带 rs641738 的人肝细胞中 CYP2C8 的活性存在显著差异（ = 0.004）。我们还观察到，携带 rs2642438 的人肝细胞中 CYP2C9 的活性在次要纯合子中显著低于非次要纯合子（ = 0.001）。在携带 rs738409、rs641738、GCKR rs780094、rs72613567 和 rs2642438 遗传变异的人肝细胞中，CYP2E1、CYP2C8、CYP2D6、CYP2E1、CYP3A4、ECOD、FMO、MAO、AO 和 CES2 的活性以及任何 II 相肝酶的活性均无显著差异。这些发现初步评估了遗传变异对健康人肝细胞中药物代谢细胞色素 P450（CYP）酶的影响，这可能对未来的药物发现研究有用。
