title: 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: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Joseph Ignatius
authors: JI
authors: Irudayam
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Deisy
authors: D
authors: Contreras
authors: Cedars-Sinai Genomics Core, Medical Genetics Institute, Cedars-Sinai Medical Center Los Angeles, CA 90048, USA.
authors: Lindsay
authors: L
authors: Spurka
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Aparna
authors: A
authors: Subramanian
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Jenieke
authors: J
authors: Allen
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Songyang
authors: S
authors: Ren
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors: Vidhya
authors: V
authors: Kanagavel
authors: Cedars-Sinai Genomics Core, Medical Genetics Institute, Cedars-Sinai Medical Center Los Angeles, CA 90048, USA.
authors: Quoclinh
authors: Q
authors: Nguyen
authors: Centre for Infectious Disease Research, Indian Institute of Science, Bangalore, Karnataka 560012, India.|Hindustan Genomics Institute, SVA Medical Center, Kadayam, Tamil Nadu 627415, India.
authors: Arunachalam
authors: A
authors: Ramaiah
authors: Department of Biotechnology, Manonmaniam Sundaranar University, Tirunelveli, Tamil Nadu 627012, India.|Hindustan Genomics Institute, SVA Medical Center, Kadayam, Tamil Nadu 627415, India.
authors: Kalidas
authors: K
authors: Ramamoorthy
authors: Department of Pathology and Laboratory Medicine, University of California at Los Angeles, Los Angeles CA 90095, USA.
authors: Samuel W
authors: SW
authors: French
authors: Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.|Department of Surgery, University of California at Los Angeles, Los Angeles CA 90095, USA.
authors: Andrew S
authors: AS
authors: Klein
authors: Cedars-Sinai Genomics Core, Medical Genetics Institute, Cedars-Sinai Medical Center Los Angeles, CA 90048, USA.
authors: Vincent
authors: V
authors: Funari
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.|Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.|Department of Surgery, University of California at Los Angeles, Los Angeles CA 90095, USA.
authors: Vaithilingaraja
authors: V
authors: Arumugaswami
mesh_terms: D053299:Apolipoprotein B-100 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007370:Interferon Type I / Q000235:genetics / Q000378:metabolism*; D016898:Interferon-alpha / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017423:Sequence Analysis, RNA; D059467:Transcriptome / Q000187:drug effects; D014779:Virus Replication; D020663:ras Guanine Nucleotide Exchange Factors
chemical_list: D053299:Apolipoprotein B-100; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016207:Cytokines; D020662:Guanine Nucleotide Exchange Factors; D007370:Interferon Type I; D016898:Interferon-alpha; C413212:RASGRP3 protein, human; C112841:TRANK1 protein, human; D020663:ras Guanine Nucleotide Exchange Factors; D012313:RNA
keywords: Differentiation; Endoderm; Hepatocyte-like cells; Hepatocytes; ISG; Innate immunity; Interferon; Interferon-stimulated genes; Pluripotent stem cells