title: Unfolded protein response is required for the definitive endodermal specification of mouse embryonic stem cells via Smad2 and β-catenin signaling.
abstract: Tremendous efforts have been made to elucidate the molecular mechanisms that control the specification of definitive endoderm cell fate in gene knockout mouse models and ES cell (ESC) differentiation models. However, the impact of the unfolded protein response (UPR), because of the stress of the endoplasmic reticulum on endodermal specification, is not well addressed. We employed UPR-inducing agents, thapsigargin and tunicamycin, in vitro to induce endodermal differentiation of mouse ESCs. Apart from the endodermal specification of ESCs, Western blotting demonstrated the enhanced phosphorylation of Smad2 and nuclear translocation of β-catenin in ESC-derived cells. The inclusion of the endoplasmic reticulum stress inhibitor tauroursodeoxycholic acid to the induction cultures prevented the differentiation of ESCs into definitive endodermal cells even when Activin A was supplemented. Also, the addition of the TGF-β inhibitor SB431542 and the Wnt/β-catenin antagonist IWP-2 negated the endodermal differentiation of ESCs mediated by thapsigargin and tunicamycin. These data suggest that the activation of the UPR appears to orchestrate the induction of the definitive endodermal cell fate of ESCs via both the Smad2 and β-catenin signaling pathways. The prospective regulatory machinery may be helpful for directing ESCs to differentiate into definitive endodermal cells for cellular therapy in the future.
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine and Shanghai Jiao Tong University, Shanghai 200127, China; School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200127, China and.
authors.forename: Huiming
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China.
authors.forename: Kam Sze
authors.initials: KS
authors.lastname: Tsang
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine and Shanghai Jiao Tong University, Shanghai 200127, China; School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200127, China and.
authors.forename: Yonghui
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Medicine and Therapeutics, The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China; Li Ka Shing Institute of Health Sciences and The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China.
authors.forename: Juliana Cn
authors.initials: JC
authors.lastname: Chan
authors.affiliation: Department of Medicine and Therapeutics, The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China; Shenzhen Research Institute, The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China; Li Ka Shing Institute of Health Sciences and The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong, China. Electronic address: gangxu@cuhk.edu.hk.
authors.forename: Gang
authors.initials: G
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine and Shanghai Jiao Tong University, Shanghai 200127, China; School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200127, China and. Electronic address: gao.weiqiang@sjtu.edu.cn.
authors.forename: Wei-Qiang
authors.initials: WQ
authors.lastname: Gao
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000502:physiology*; D004707:Endoderm / Q000166:cytology; D059865:Endoplasmic Reticulum Stress; D051379:Mice; D051899:Smad2 Protein / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000502:physiology; D014415:Tunicamycin / Q000494:pharmacology; D056811:Unfolded Protein Response; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism*
chemical_list: C495267:CTNNB1 protein, mouse; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D016212:Transforming Growth Factor beta; D051176:beta Catenin; D014415:Tunicamycin; D019284:Thapsigargin
keywords: Cell Culture; Cell Signaling; Development; Differentiation; Embryonic Stem Cell; Endoplasmic Reticulum Stress (ER Stress)
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