title: Induction of LYVE-1/stabilin-2-positive liver sinusoidal endothelial-like cells from embryoid bodies by modulation of adrenomedullin-RAMP2 signaling.
abstract: Embryonic stem cells (ESCs) are a useful source for various cell lineages. So far, however, progress toward reconstitution of mature liver morphology and function has been limited. We have shown that knockout mice deficient in adrenomedullin (AM), a multifunctional endogenous peptide, or its receptor-activity modifying protein (RAMP2) die in utero due to poor vascular development and hemorrhage within the liver. In this study, using embryoid bodies (EBs)-culture system, we successfully induced liver sinusoidal endothelial-like cells by modulation of AM-RAMP2. In an EB differentiation system, we found that co-administration of AM and SB431542, an inhibitor of transforming growth factor β (TGFβ) receptor type 1, markedly enhanced differentiation of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1)/stabilin-2-positive endothelial cells. These cells showed robust endocytosis of acetylated low-density lipoprotein (Ac-LDL) and upregulated expression of liver sinusoidal endothelial cells (LSECs)-specific markers, including factor 8 (F8), Fc-γ receptor 2b (Fcgr2b), and mannose receptor C type 1 (Mrc1), and also possessed fenestrae-like structure, a key morphological feature of LSECs. In RAMP2-null liver, by contrast, LYVE-1 was downregulated in LSECs, and the sinusoidal structure was disrupted. Our findings highlight the importance of AM-RAMP2 signaling for development of LSECs.
authors.affiliation: Department of Organ Regeneration, Shinshu University Graduate School of Medicine, Asahi 3-1-1, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Arai
authors.forename: Takayuki
authors.initials: T
authors.lastname: Sakurai
authors.forename: Akiko
authors.initials: A
authors.lastname: Kamiyoshi
authors.forename: Yuka
authors.initials: Y
authors.lastname: Ichikawa-Shindo
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Iinuma
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Iesato
authors.forename: Teruhide
authors.initials: T
authors.lastname: Koyama
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yoshizawa
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Uetake
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yamauchi
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
authors.forename: Hisaka
authors.initials: H
authors.lastname: Kawate
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ogawa
authors.forename: Akira
authors.initials: A
authors.lastname: Kobayashi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Takayuki
authors.initials: T
authors.lastname: Shindo
mesh_terms: D053607:Adrenomedullin / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D004149:Dioxoles / Q000494:pharmacology; D004357:Drug Synergism; D053595:Embryonic Stem Cells / Q000166:cytology; D004705:Endocytosis; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D018507:Gene Expression Regulation, Developmental; D006023:Glycoproteins / Q000378:metabolism*; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology; D026901:Membrane Transport Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning; D009024:Morphogenesis; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D058263:Receptor Activity-Modifying Protein 2 / Q000235:genetics / Q000378:metabolism*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D015398:Signal Transduction
chemical_list: D053607:Adrenomedullin; D001549:Benzamides; D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal; D004149:Dioxoles; D006023:Glycoproteins; D026901:Membrane Transport Proteins; D017346:Protein Serine-Threonine Kinases; D058263:Receptor Activity-Modifying Protein 2; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta; C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; C545506:Ramp2 protein, mouse; C486733:Stab2 protein, mouse; C492273:LYVE1 protein, mouse