title: Connexin 32-mediated cell-cell communication is essential for hepatic differentiation from human embryonic stem cells.
abstract: Gap junction-mediated cell-cell interactions are highly conserved and play essential roles in cell survival, proliferation, differentiation and patterning. We report that Connexin 32 (Cx32)-mediated gap junctional intercellular communication (GJIC) is necessary for human embryonic stem cell-derived hepatocytes (hESC-Heps) during step-wise hepatic lineage restriction and maturation. Vitamin K2, previously shown to promote Cx32 expression in mature hepatocytes, up-regulated Cx32 expression and GJIC activation during hepatic differentiation and maturation, resulting in significant increases of hepatic markers expression and hepatocyte functions. In contrast, negative Cx32 regulator 2-aminoethoxydiphenyl borate blocked hESC-to-hepatocyte maturation and muted hepatocyte functions through disruption of GJIC activities. Dynamic gap junction organization and internalization are phosphorylation-dependent and the p38 mitogen-activated protein kinases pathway (MAPK) can negatively regulate Cxs through phosphorylation-dependent degradation of Cxs. We found that p38 MAPK inhibitor SB203580 improved maturation of hESC-Heps correlating with up-regulation of Cx32; by contrast, the p38 MAPK activator, anisomycin, blocked hESC-Heps maturation correlating with down-regulation of Cx32. These results suggested that Cx32 is essential for cell-cell interactions that facilitate driving hESCs through hepatic-lineage maturation. Regulators of both Cx32 and other members of its pathways maybe used as a promising approach on regulating hepatic lineage restriction of pluripotent stem cells and optimizing their functional maturation.
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|South China Research Center for Stem Cell and Regenerative Medicine, South China Institute of Biomedicine, Guangzhou 510005, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Qin
authors.affiliation: Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Mingyang
authors.initials: M
authors.lastname: Chang
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|South China Research Center for Stem Cell and Regenerative Medicine, South China Institute of Biomedicine, Guangzhou 510005, China.
authors.forename: Shuyong
authors.initials: S
authors.lastname: Wang
authors.affiliation: CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Zhenbo
authors.initials: Z
authors.lastname: Liu
authors.affiliation: CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.affiliation: Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Yan
authors.affiliation: Laboratory of Hematological Pharmacology, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|South China Research Center for Stem Cell and Regenerative Medicine, South China Institute of Biomedicine, Guangzhou 510005, China.
authors.forename: Bowen
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Laboratory of Hematological Pharmacology, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Guifang
authors.initials: G
authors.lastname: Dou
authors.affiliation: CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Liu
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.|South China Research Center for Stem Cell and Regenerative Medicine, South China Institute of Biomedicine, Guangzhou 510005, China.
authors.forename: Xuetao
authors.initials: X
authors.lastname: Pei
authors.affiliation: Tissue Engineering Lab, Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Yunfang
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000841:Anisomycin / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D001896:Boron Compounds / Q000494:pharmacology; D002450:Cell Communication; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D017630:Connexins / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D008099:Liver / Q000166:cytology*; D010766:Phosphorylation / Q000187:drug effects; D011725:Pyridines / Q000494:pharmacology; D024482:Vitamin K 2 / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D000097002:Gap Junction beta-1 Protein
chemical_list: D000841:Anisomycin; D015415:Biomarkers; D001896:Boron Compounds; D017630:Connexins; D003577:Cytochrome P-450 Enzyme System; D007093:Imidazoles; D011725:Pyridines; D024482:Vitamin K 2; D048051:p38 Mitogen-Activated Protein Kinases; D000097002:Gap Junction beta-1 Protein; C109986:2-aminoethoxydiphenyl borate; C093642:SB 203580