Characterization of type I interferon pathway during hepatic differentiation of human pluripotent stem cells and hepatitis C virus infection.

Abstract

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 (hESCs). 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.

• Authors: Joseph Ignatius Irudayam, Deisy Contreras, Lindsay Spurka, Aparna Subramanian, Jenieke Allen, Songyang Ren, Vidhya Kanagavel, Quoclinh Nguyen, Arunachalam Ramaiah, Kalidas Ramamoorthy, Samuel W French, Andrew S Klein, Vincent Funari, Vaithilingaraja Arumugaswami
• Journal: Stem cell research (Stem Cell Res) Vol. 15 Issue 2 Pg. 354-364 (Sep 2015) ISSN: 1876-7753 [Electronic] England
• PMID: 26313525 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015. Published by Elsevier B.V.

Chemical References

• Apolipoprotein B-100
• CLMP protein, human
• Coxsackie and Adenovirus Receptor-Like Membrane Protein
• Cytokines
• Guanine Nucleotide Exchange Factors
• Interferon Type I
• Interferon-alpha
• RASGRP3 protein, human
• TRANK1 protein, human
• ras Guanine Nucleotide Exchange Factors
• RNA

Topics

• Apolipoprotein B-100 (metabolism)
• Cell Differentiation (drug effects)
• Cell Line
• Cell Lineage
• Coxsackie and Adenovirus Receptor-Like Membrane Protein (genetics, metabolism)
• Cytokines (genetics, metabolism)
• Gene Expression Profiling
• Guanine Nucleotide Exchange Factors (genetics, metabolism)
• Hepacivirus (genetics, physiology)
• Hepatocytes (cytology, metabolism)
• Humans
• Interferon Type I (genetics, metabolism)
• Interferon-alpha (pharmacology)
• Liver (metabolism)
• Pluripotent Stem Cells (cytology, metabolism)
• RNA (chemistry, isolation & purification, metabolism)
• Sequence Analysis, RNA
• Transcriptome (drug effects)
• Virus Replication
• ras Guanine Nucleotide Exchange Factors