Characterization of type I interferon pathway during hepatic differentiation of human pluripotent stem cells and hepatitis C virus infection.
Characterization of type I interferon pathway during hepatic differentiation of human pluripotent stem cells and hepatitis C virus infection.
复制标题
DOI：
10.1016/j.scr.2015.08.003
复制
发表时间：
2015-09
影响因子：
1.2
通讯作者：
Arumugaswami V
中科院分区：
文献类型：
--
作者：
Irudayam JI;Contreras D;Spurka L;Subramanian A;Allen J;Ren S;Kanagavel V;Nguyen Q;Ramaiah A;Ramamoorthy K;French SW;Klein AS;Funari V;Arumugaswami V
人们正在积极研究多能干细胞作为再生受损肝脏的细胞来源。为了使移植细胞在体内长期存活，细胞不仅必须执行肝脏特有的功能，而且还要承受物理压力和入侵的病原体。由干扰素 (IFN) 途径协调的细胞先天免疫系统提供了抵御病原体的第一道防线。本研究的目的是评估先天免疫功能，并系统地分析多能干细胞肝分化过程中 IFN 诱导的基因。为了实现这一目标，我们从人胚胎干细胞 (hESC) 中衍生出内胚层细胞（分化后第 5 天）、成肝细胞（第 15 天）和肝细胞样细胞（第 21 天）。第5、15和21天的细胞用IFN-α刺激并进行IFN途径分析。通过RNA测序进行转录组分析。结果表明，IFN-α处理激活了分化细胞中的STAT-JAK通路。转录组分析表明经典和非经典 IFN 刺激基因 (ISG) 的阶段特异性表达。随后的验证证实了 IFN 处理后分化的肝细胞表达了新型 ISG，包括 RASGRP3、CLMP 和 TRANK1。丙型肝炎病毒在 hESC 衍生的肝细胞中复制诱导 ISG 的表达 - LAMP3、ETV7、RASGRP3 和 TRANK1。 hESC 衍生的肝细胞含有完整的先天系统，可以识别入侵的病原体。除了评估细胞治疗应用的组织特异性功能外，测试移植细胞的先天免疫功能也很重要，以确保充分防御感染并提高移植物存活率。
Pluripotent stem cells are being actively studied as a cell source for regenerating damaged liver. For long term survival of engrafting cells in the body, not only do the cells have to execute liver-specific function but also withstand the physical strains and invading pathogens. The cellular innate immune system orchestrated by the interferon (IFN) pathway provides the first line of defense against pathogens. The objective of this study is to assess the innate immune function as well as to systematically profile the IFN-induced genes during hepatic differentiation of pluripotent stem cells. To address this objective, we derived endodermal cells (day 5 post-differentiation), hepatoblast (day 15) and hepatocyte-like cells (day 21) from human embryonic stem cells (hESC). Day 5, 15 and 21 cells were stimulated with IFN-α and subjected to IFN pathway analysis. Transcriptome analysis was carried out by RNA sequencing. The results showed that the IFN-α treatment activated STAT-JAK pathway in differentiating cells. Transcriptome analysis indicated stage specific expression of classical and non-classical IFN-stimulated genes (ISGs). Subsequent validation confirmed the expression of novel ISGs including RASGRP3, CLMP and TRANK1 by differentiated hepatic cells upon IFN treatment. Hepatitis C virus replication in hESC-derived hepatic cells induced the expression of ISGs – LAMP3, ETV7, RASGRP3, and TRANK1. The hESC-derived hepatic cells contain intact innate system and can recognize invading pathogens. Besides assessing the tissue-specific functions for cell therapy applications, it may also be important to test the innate immune function of engrafting cells to ensure adequate defense against infections and improve graft survival.