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GIPC-Regulated IGFBP-3 Promotes HSC Migration In Vitro and Portal Hypertension In Vivo Through a β1-Integrin Pathway.
Cellular and Molecular Gastroenterology and Hepatology ( IF 7.2 ) Pub Date : 2020-05-22 , DOI: 10.1016/j.jcmgh.2020.05.005
Usman Yaqoob 1 , Fanghong Luo 2 , Thomas Greuter 3 , Nidhi Jalan Sakrikar 1 , Tejasav S Sehrawat 1 , Jianwen Lu 1 , Xiao Hu 1 , Jinhang Gao 1 , Enis Kostallari 1 , Jingbiao Chen 1 , Juan Pablo Arab 1 , Rosa Martin-Mateos 4 , Sheng Cao 1 , Vijay H Shah 1
Affiliation  

Background & Aims

Transforming growth factor (TGF-β)–induced activation of quiescent hepatic stellate cells (HSCs) and their transformation to myofibroblasts is a key event in liver fibrosis and portal hypertension. GIPC (also referred to as synectin) is a downstream signal activation molecule of TGF-β and other receptors. In this study, we sought to identify novel genes targeted by TGF-β and GIPC and elucidate if and how they may contribute to liver fibrosis.

Methods

We performed sequential messenger RNA sequencing analysis on TGF-β–stimulated HSCs and then on TGF-β–stimulated HSCs in the presence and absence of GIPC also referred to as synectin (GIPC) knockdown. Insulin-like growth factor binding protein-3 (IGFBP-3) transport protein emerged as a top activation target of both TGF-β and GIPC. Quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, targeted chromatin immunoprecipitation, and Western blot analysis were done for further confirmation.

Results

IGFBP-3, an insulin growth factor transport protein, emerged as a top activation target of both TGF-β and GIPC, which was confirmed by quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot analysis. Targeted chromatin immunoprecipitation showed that GIPC increases the histone 3 lysine 27 (H3K27) acetylation activating mark and concurrently decreases the H3K27 inhibitory trimethylation (H3K27m3) mark, providing an epigenetic correlate to the gene regulation changes. In vivo, global knockout of IGFBP-3 mice resulted in attenuation of HSC activation markers and attenuation of portal pressure in response to chronic liver injury models. Analysis of serum levels from cirrhotic patients also showed an IGFBP-3 increase of more than 2-fold compared with healthy controls. Finally, in vitro mechanism studies showed that IGFBP-3 promotes HSC migration through integrin-dependent phosphorylation of protein kinase B.

Conclusions

TGF-β up-regulates IGFBP-3 through GIPC, leading to increased HSC migration in vitro and promotes portal hypertension in vivo. These studies support the role of IGFBP-3 as a potential pathophysiologic target or biomarker in chronic liver disease.



中文翻译:

GIPC 调节的 IGFBP-3 通过 β1-整合素途径促进体外 HSC 迁移和体内门静脉高压。

背景与目标

转化生长因子 (TGF-β) 诱导的静止肝星状细胞 (HSC) 激活及其向肌成纤维细胞的转化是肝纤维化和门静脉高压症的关键事件。GIPC(也称为synectin)是TGF-β和其他受体的下游信号激活分子。在这项研究中,我们试图确定 TGF-β 和 GIPC 靶向的新基因,并阐明它们是否以及如何导致肝纤维化。

方法

我们对 TGF-β 刺激的 HSCs 进行了序列信使 RNA 测序分析,然后在存在和不存在 GIPC 的情况下对 TGF-β 刺激的 HSCs 进行了序列分析,也称为连接蛋白 (GIPC) 敲低。胰岛素样生长因子结合蛋白-3 (IGFBP-3) 转运蛋白成为 TGF-β 和 GIPC 的首要激活靶标。进行定量聚合酶链反应、酶联免疫吸附测定、靶向染色质免疫沉淀和蛋白质印迹分析以进一步确认。

结果

IGFBP-3 是一种胰岛素生长因子转运蛋白,是 TGF-β 和 GIPC 的首要激活靶标,这一点已通过定量聚合酶链反应、酶联免疫吸附测定和蛋白质印迹分析得到证实。靶向染色质免疫沉淀显示 GIPC 增加组蛋白 3 赖氨酸 27 (H3K27) 乙酰化激活标记并同时降低 H3K27 抑制性三甲基化 (H3K27m3) 标记,提供与基因调控变化相关的表观遗传学。在体内,IGFBP-3 小鼠的整体敲除导致 HSC 激活标志物的减弱和门静脉压力的减弱,以响应慢性肝损伤模型。肝硬化患者的血清水平分析也显示,与健康对照相比,IGFBP-3 增加了 2 倍以上。最后,

结论

TGF-β 通过 GIPC 上调 IGFBP-3,导致体外 HSC 迁移增加并促进体内门静脉高压。这些研究支持 IGFBP-3 作为慢性肝病潜在病理生理靶点或生物标志物的作用。

更新日期:2020-05-22
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