Stabilization of HIF through inhibition of Cullin-2 neddylation is protective in mucosal inflammatory responses
Stabilization of HIF through inhibition of Cullin-2 neddylation is protective in mucosal inflammatory responses
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DOI：
10.1096/fj.14-259663
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发表时间：
2015-01-01
期刊：
影响因子：
4.8
通讯作者：
Colgan, Sean P.
中科院分区：
文献类型：
--
作者：
Curtis, Valerie F.;Ehrentraut, Stefan F.;Colgan, Sean P.
人们对理解炎症性疾病中蛋白质的翻译后修饰很感兴趣。Neddylation是神经前体细胞表达的分子NEDD 8（发育下调8）的缀合，以促进蛋白质稳定。Cullin是NEDD 8靶点家族，在蛋白质的稳定和降解中起重要作用，如缺氧诱导因子（HIF;通过Cullin-2）。在这里，我们阐明的作用，人deneddylase-1（DEN-1，也称为SENP 8）在体外和体内的炎症反应，并定义条件靶向neddylation在粘膜炎症模型。HIF在炎症模型中提供保护，因此我们检查了DEN-1对HIF稳定的贡献。MLN 4924（IC 50，4.7 nM）对neddylation活性的药理学靶向作用可稳定HIF-1 α，激活HIF启动子活性2.5倍，并诱导人上皮细胞中HIF靶基因高达5倍。人肠上皮细胞中DEN-1的敲低导致屏障形成的动力学增加，通透性降低，屏障恢复增强2 +/- 0.5倍。体内平行研究显示，MLN 4924可减轻小鼠葡聚糖硫酸钠结肠炎的疾病严重程度，包括体重减轻、结肠长度和组织学严重程度。我们的结论是，DEN-1是一个调节器的cullin neddylation和微调炎症反应在体外和体内。对cullin neddylation的药理学抑制可能为粘膜炎性疾病提供治疗机会。
There is interest in understanding post-translational modifications of proteins in inflammatory disease. Neddylation is the conjugation of the molecule neural precursor cell expressed, developmentally down-regulated 8 (NEDD8) to promote protein stabilization. Cullins are a family of NEDD8 targets important in the stabilization and degradation of proteins, such as hypoxia-inducible factor (HIF; via Cullin-2). Here, we elucidate the role of human deneddylase-1 (DEN-1, also called SENP8) in inflammatory responses in vitro and in vivo and define conditions for targeting neddylation in models of mucosal inflammation. HIF provides protection in inflammatory models, so we examined the contribution of DEN-1 to HIF stabilization. Pharmacologic targeting of neddylation activity with MLN4924 (IC50, 4.7 nM) stabilized HIF-1 alpha, activated HIF promoter activity by 2.5-fold, and induced HIF-target genes in human epithelial cells up to 5-fold. Knockdown of DEN-1 in human intestinal epithelial cells resulted in increased kinetics in barrier formation, decreased permeability, and enhanced barrier restitution by 2 +/- 0.5-fold. Parallel studies in vivo revealed that MLN4924 abrogated disease severity in murine dextran sulfate sodium colitis, including weight loss, colon length, and histologic severity. We conclude that DEN-1 is a regulator of cullin neddylation and fine-tunes the inflammatory response in vitro and in vivo. Pharmacologic inhibition of cullin neddylation may provide a therapeutic opportunity in mucosal inflammatory disease.