人iPSC源性肝细胞抗体药物肝毒性评价研究

Evaluation of hepatotoxicity for antibody drugs using human iPSC-derived hepatocytes

目的 用人诱导多能干细胞(induced pluripotent stem cells,iPSC)源性肝细胞评估抗体药物的肝细胞毒性。方法 qPCR、Western blot、细胞免疫荧光检测肝细胞特征性表型分子白蛋白、尿素氮及药物代谢酶的表达;MTT法检测肝细胞生长情况;ELISA检测肝细胞白蛋白、尿素氮分泌水平;过碘酸-雪夫染色、油红O染色检测肝细胞储存糖原、脂滴的能力。结果 HcHAb18、Hab18、5A12单抗处理后人iPSC源性肝细胞生长和分泌功能并未发生明显改变;然而,肝细胞特征性表型分子表达降低,糖原和脂滴储存能力抑制,药物代谢酶表达相应改变。结论 3株单抗虽不影响人iPSC源性肝细胞的生存,但改变其表型和代谢;人iPSC源性肝细胞可用于抗体药物的肝细胞毒性评估。

Objective Human induced pluripotent stem cells(iPSC)-derived hepatocytes were utilized to evaluate the hepatotoxicity of antibody drugs.Methods qPCR,western blot and cellular immunofluorescence were used to investigate the expression of albumin,urea nitrogen and drug metabolism enzymes in hepatocytes.MTT assay was used to analyze the cellular growth hepatocyte.ELISA was used to detect the secretion function of albumin and urea nitrogen in hepatocytes.Periodic acid-Schiff and oil red O staining were used to explore the ability of hepatocytes to store glycogen and lipid droplets.Results The growth and secretion function of iPSC-derived hepatocytes did not change significantly after HcHAb18,Hab18 and 5A12 monoclonal antibody treatment;however,the molecular expression of characteristic phenotype in hepatocytes was decreased,the storage capacity of glycogen and lipid droplet was inhibited,and the expression of drug-metabolizing enzymes was correspondingly altered.Conclusion Three monoclonal antibodies do not affect the survival of human iPSC-derived hepatocytes,but change their phenotype and metabolism.Human iPSC-derived hepatocytes can be used to assess the hepatocytotoxicity of antibody drugs.