Induction of LYVE-1/stabilin-2-positive liver sinusoidal endothelial-like cells from embryoid bodies by modulation of adrenomedullin-RAMP2 signaling
Induction of LYVE-1/stabilin-2-positive liver sinusoidal endothelial-like cells from embryoid bodies by modulation of adrenomedullin-RAMP2 signaling
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DOI：
10.1016/j.peptides.2011.07.005
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发表时间：
2011-09-01
期刊：
影响因子：
3
通讯作者：
Shindo, Takayuki
中科院分区：
文献类型：
--
作者：
Arai, Takuma;Sakurai, Takayuki;Shindo, Takayuki
胚胎干细胞（ESC）是多种细胞谱系的有用来源。然而，到目前为止，成熟肝脏形态和功能重建的进展有限。我们已经证明，肾上腺髓质素（AM），一种多功能内源性肽，或其受体活性修饰蛋白（RAMP 2）基因敲除小鼠在子宫内死亡，由于不良的血管发育和肝脏内出血。在本研究中，我们利用胚状体（EBs）培养系统，通过AM-RAMP 2的调控，成功地诱导了肝窦内皮样细胞。在EB分化系统中，我们发现AM和SB 431542（一种转化生长因子β（TGF β）受体1型抑制剂）的联合给药显著增强了淋巴管内皮透明质酸受体-1（LYVE-1）/稳定素-2阳性内皮细胞的分化。这些细胞显示出乙酰化低密度脂蛋白（Ac-LDL）的稳健内吞作用和肝窦内皮细胞（LSEC）特异性标志物（包括因子8（F8）、Fc-γ受体2b（Fcgr 2b）和甘露糖受体C 1型（Mrc 1））的表达上调，并且还具有窗孔样结构（LSEC的关键形态学特征）。相反，在RAMP 2缺失的肝脏中，LYVE-1在LSEC中下调，并且窦状结构被破坏。我们的研究结果强调了AM-RAMP 2信号传导对LSEC发展的重要性。(C)2011 Elsevier Inc. All rights reserved.
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 beta (TGF beta) 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-gamma 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. (C) 2011 Elsevier Inc. All rights reserved.