Induction of LYVE-1/stabilin-2-positive liver sinusoidal endothelial-like cells from embryoid bodies by modulation of adrenomedullin-RAMP2 signaling

• Published in: Peptides (New York, N.Y. : 1980) N.Y. : 1980), 2011-09, Vol.32 (9), p.1855-1865
• Main Authors: Arai, Takuma, Sakurai, Takayuki, Kamiyoshi, Akiko, Ichikawa-Shindo, Yuka, Iinuma, Nobuyoshi, Iesato, Yasuhiro, Koyama, Teruhide, Yoshizawa, Takahiro, Uetake, Ryuichi, Yamauchi, Akihiro, Yang, Lei, Kawate, Hisaka, Ogawa, Shinichiro, Kobayashi, Akira, Miyagawa, Shinichi, Shindo, Takayuki
• Format: Article
• Language: English
• Subjects: Adrenomedullin (AM); Adrenomedullin - metabolism; Adrenomedullin - pharmacology; Animals; Benzamides - pharmacology; Biological and medical sciences; Biomarkers; Cell Adhesion Molecules, Neuronal - metabolism; Cell Differentiation; Cells, Cultured; Dioxoles - pharmacology; Drug Synergism; Embryonic stem cells; Embryonic Stem Cells - cytology; Endocytosis; endothelial cells; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelial Cells - ultrastructure; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Developmental; Glycoproteins - metabolism; hemorrhage; hyaluronic acid; Immunohistochemistry; knockout mutants; liver; Liver - cytology; Liver sinusoidal endothelial cells (LSEC); low density lipoprotein; Lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1); mannose; Mice; Mice, Inbred C57BL; Microscopy, Electron, Scanning; Morphogenesis; Phenotype; Protein-Serine-Threonine Kinases - antagonists & inhibitors; Receptor activity-modifying protein (RAMP); Receptor Activity-Modifying Protein 2 - genetics; Receptor Activity-Modifying Protein 2 - metabolism; Receptors, Transforming Growth Factor beta - antagonists & inhibitors; Signal Transduction; Stabilin-2; transforming growth factor beta; Vertebrates: endocrinology
• ISSN: 0196-9781; 1873-5169
• DOI: 10.1016/j.peptides.2011.07.005

► Co-administration of AM and SB431542 enhanced differentiation of LYVE-1/stabilin-2-positive endothelial cells in EBs-culture system. ► These cells showed robust endocytosis and upregulated expression of LSEC-specific markers and possess fenestrae-like structure. ► In RAMP2-null liver, LYVE-1 was downregulated in LSEC, and the sinusoidal structure was disrupted. ► Our findings highlight the importance of AM–RAMP2 signaling for development of LSEC. 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.