Modeling Human Bile Acid Transport and Synthesis in Stem Cell-Derived Hepatocytes with a Patient-Specific Mutation.
Modeling Human Bile Acid Transport and Synthesis in Stem Cell-Derived Hepatocytes with a Patient-Specific Mutation.
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DOI：
10.1016/j.stemcr.2020.12.008
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发表时间：
2021-02-09
影响因子：
5.9
通讯作者：
Asai A
中科院分区：
文献类型：
--
作者：
Hayashi H;Osaka S;Sakabe K;Fukami A;Kishimoto E;Aihara E;Sabu Y;Mizutani A;Kusuhara H;Naritaka N;Zhang W;Huppert SS;Sakabe M;Nakamura T;Hu YC;Mayhew C;Setchell K;Takebe T;Asai A
胆盐输出泵（BSEP）负责从肝细胞输出胆汁酸。BSEP突变的肝细胞中胆汁酸跨细胞转运受损导致胆汁淤积。在BSEP缺乏的人肝细胞中调节细胞内胆汁酸浓度的代偿机制尚不清楚。为了确定防止胆汁酸在肝细胞中细胞毒性蓄积的途径，我们在Transwell培养系统中开发了基于人诱导多能干细胞的同基因BSEP缺陷肝细胞模型。诱导的肝细胞（i-Heps）表现出缺陷的顶端出口的胆汁酸，但保持低细胞内胆汁酸浓度诱导基底外侧出口。模拟胆汁酸对肝细胞的自动调节，我们发现BSEP缺陷的i-Heps通过基底外侧摄取和输出而无顶端输出使用FXR途径抑制从头胆汁酸合成。这些观察结果为开发治疗靶点提供了信息，以减少BSEP缺乏症患者的总体胆汁酸池。通过CRISPR产生人同基因iPSC以研究BSEP iPSC衍生的肝细胞的截短突变重现患者中BSEP缺乏的病理生理学BSEP缺乏的肝细胞诱导替代基底外侧胆汁酸输出FXR的激活抑制BSEP缺乏的肝细胞中的从头胆汁酸合成在该研究中，Asai及其同事为胆汁酸转运缺陷引起的人类遗传性肝病提供了新疾病模型的概念证明。同基因iPSC衍生的肝细胞中的胆盐输出泵（BSEP）突变重现了疾病表型。这种新的模型揭示了BSEP缺陷肝细胞中的适应性反向胆汁酸输出，同时保持从头胆汁酸合成的调节反馈。
The bile salt export pump (BSEP) is responsible for the export of bile acid from hepatocytes. Impaired transcellular transport of bile acids in hepatocytes with mutations in BSEP causes cholestasis. Compensatory mechanisms to regulate the intracellular bile acid concentration in human hepatocytes with BSEP deficiency remain unclear. To define pathways that prevent cytotoxic accumulation of bile acid in hepatocytes, we developed a human induced pluripotent stem cell-based model of isogenic BSEP-deficient hepatocytes in a Transwell culture system. Induced hepatocytes (i-Heps) exhibited defects in the apical export of bile acids but maintained a low intracellular bile acid concentration by inducing basolateral export. Modeling the autoregulation of bile acids on hepatocytes, we found that BSEP-deficient i-Heps suppressed de novo bile acid synthesis using the FXR pathway via basolateral uptake and export without apical export. These observations inform the development of therapeutic targets to reduce the overall bile acid pool in patients with BSEP deficiency. Human isogenic iPSCs were generated by CRISPR to study a truncating mutation of BSEP iPSC-derived hepatocytes recapitulate pathophysiology of BSEP deficiency in patients BSEP-deficient hepatocytes induce alternative basolateral bile acid export Activation of FXR suppresses de novo bile acid synthesis in BSEP-deficient hepatocytes In this study, Asai and colleagues provide proof of concept of novel disease modeling for human genetic liver disorders caused by bile acid transport defects. A bile salt export pump (BSEP) mutation in isogenic iPSC-derived hepatocytes recapitulated the disease phenotype. This novel model revealed adaptive reverse bile acid exports in BSEP-deficient hepatocytes while maintaining regulatory feedback of de novo bile acid synthesis.
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影响因子：
10.6
作者：
Ballatori, Nazzareno;Christian, Whitney V.;Wheeler, Sadie G.;Hammond, Christine L.
通讯作者：
Hammond, Christine L.
影响因子：
5.8
作者：
Cheung, Leanna;Flemming, Claudia L.;Norris, Murray D.
通讯作者：
Norris, Murray D.
影响因子：
13.5
作者：
Wang, RX;Lam, P;Ling, V
通讯作者：
Ling, V
影响因子：
4.8
作者：
Christian, Whitney V.;Li, Na;Ballatori, Nazzareno
通讯作者：
Ballatori, Nazzareno
溶质载体有机阴离子转运蛋白家族成员 3A1 是胆汁酸流出转运蛋白在胆汁淤积中的作用
影响因子：
29.4
作者：
Pan Q;Zhang X;Zhang L;Cheng Y;Zhao N;Li F;Zhou X;Chen S;Li J;Xu S;Huang D;Chen Y;Li L;Wang H;Chen W;Cai SY;Boyer JL;Chai J
通讯作者：
Chai J