An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells
An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells
复制标题
DOI：
10.1016/j.scr.2018.08.018
复制
发表时间：
2018-10-01
影响因子：
1.2
通讯作者：
Inagaki, Yutaka
中科院分区：
文献类型：
--
作者：
Kamiya, Akihide;Chikada, Hiromi;Inagaki, Yutaka
在发育中的肝脏中，胆管结构是由肝祖细胞(HPC)分化为胆管细胞形成的。以胆管细胞失控扩张为特征的多囊性肝病的一种亚型，是由蛋白激酶C底物80K-H(PRKCSH)等基因异常引起的。在这项研究中，我们旨在通过编辑人类可诱导多能干细胞(IPS)中的PRKCSH基因来模拟体外的疾病过程。体外培养的人iPS细胞来源的CD13(+)CD133(+)HPC有一部分分化为CD13(-)细胞。在随后的凝胶包埋培养过程中，CD13(-)细胞形成具有上皮细胞极性的胆管标志物阳性的囊状结构。PRKCSH基因缺失增加了CD13(-)细胞中胆管细胞转录因子的表达，增加了胆管细胞囊性结构的数量。这些结果提示PRKCSH缺乏促进了HPC来源的胆管细胞的分化，为分析多囊疾病的分子机制提供了良好的体外模型。
In the developing liver, bile duct structure is formed through differentiation of hepatic progenitor cells (HPC) into cholangiocytes. A subtype of polycystic liver diseases characterized by uncontrolled expansion of bile ductal cells is caused by genetic abnormalities such as in that of protein kinase C substrate 80 K-H (PRKCSH). In this study, we aimed to mimic the disease process in vitro by genome editing of the PRKCSH locus in human inducible pluripotent stem (iPS) cells. A proportion of cultured human iPS cell-derived CD13(+) CD133(+) HPC differentiated into CD13(-) cells. During the subsequent gel embedding culture, CD13(-) cells formed bile ductal marker-positive cystic structures with the polarity of epithelial cells. A deletion of PRKCSH gene increased expression of cholangiocytic transcription factors in CD13(-) cells and the number of cholangiocytic cyst structure. These results suggest that PRKCSH deficiency promotes the differentiation of HPC-derived cholangiocytes, providing a good in vitro model to analyze the molecular mechanisms underlying polycystic diseases.