The NEDD8-activating enzyme inhibitor MLN4924 reduces ischemic brain injury in mice.
The NEDD8-activating enzyme inhibitor MLN4924 reduces ischemic brain injury in mice.
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NEDD8 激活酶抑制剂 MLN4924 可减轻小鼠缺血性脑损伤
DOI：
10.1073/pnas.2111896119
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发表时间：
2022-02-08
影响因子：
11.1
通讯作者：
Zhao BQ
中科院分区：
文献类型：
--
作者：
Yu H;Luo H;Chang L;Wang S;Geng X;Kang L;Zhong Y;Cao Y;Wang R;Yang X;Zhu Y;Shi MJ;Hu Y;Liu Z;Yin X;Ran Y;Yang H;Fan W;Zhao BQ
缺血性中风是导致死亡和残疾的主要原因，治疗方法有限。神经元前体细胞表达的发育下调蛋白8 （NEDD8）是一种泛素样蛋白，参与蛋白质类化修饰。一流的抗癌药物MLN4924由于特异性抑制nedd8活化酶，在抑制肿瘤发生和减轻炎症反应中起着至关重要的作用。在此，我们研究了MLN4924在实验性脑卒中后的潜在保护作用。我们发现，在脑缺血后，类化修饰通路在大脑中过度激活。MLN4924通过抑制中性粒细胞外渗和维持血脑屏障的完整性来保护脑免受缺血性损伤。我们的发现为抑制类化修饰治疗缺血性脑损伤提供了新的思路。血脑屏障（BBB）的破坏和发生在血脑屏障的炎症在缺血性卒中的病理生理中起着关键的，主要是有害的作用。类泛素化是一种泛素化途径，通过将神经元前体细胞表达的发育下调蛋白8 （NEDD8）偶联到靶蛋白上，在各种细胞功能中起着至关重要的作用。然而，类化修饰通路在缺血性脑卒中中的作用尚不明确。在这里，我们报道了NEDD8结合在缺血性卒中后的急性期增加，并且存在于血管内和骨髓内的中性粒细胞中。MLN4924（也称为pevonedistat）抑制类化修饰，灭活cullin-RING E3连接酶（CRL），减少脑梗死和改善功能结局。MLN4924治疗诱导了CRL底物神经纤维瘤病1 （NF1）的积累。通过使用病毒介导的NF1沉默，我们发现NF1敲除消除了mln4924依赖的中性粒细胞运输抑制。这些作用是通过内皮细胞p选择素和细胞间粘附分子-1 （ICAM-1）的激活介导的，抗p选择素或抗ICAM-1抗体的阻断抗体逆转NF1沉默诱导的mln4924处理小鼠中性粒细胞浸润的增加。此外，我们发现NF1沉默阻断mln4924通过激活缺血性卒中后脑微血管中的蛋白激酶Cδ (PKCδ)、肉myristoylated富丙氨酸C激酶底物（MARCKS）和肌球蛋白轻链（MLC）来提供血脑屏障保护和神经保护，并且用PKCδ抑制剂rottlerin治疗小鼠可降低这种增加的血脑屏障通透性。我们的研究表明，类化修饰的增加促进了中性粒细胞的运输，从而加剧了血脑屏障的损伤和卒中的结果。我们认为类化抑制可能对缺血性脑卒中有益。
Ischemic stroke is a leading cause of death and disability with limited therapies. Neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8) is a ubiquitin-like protein that is involved in protein neddylation. The first-in-class anticancer agent called MLN4924 plays a crucial role in suppressing tumorigenesis and attenuating inflammatory responses due to specifically inhibiting NEDD8-activating enzyme. Here, we investigated the potential protective role of MLN4924 after experimental stroke. We showed that the neddylation pathway is overactivated in the brain following cerebral ischemia. Inhibition of neddylation by MLN4924 protects the brain against ischemic injury by attenuating neutrophil extravasation and maintaining blood–brain barrier integrity. Our findings provide insights into the promising treatment with neddylation inhibition for ischemic brain injury. Blood–brain barrier (BBB) breakdown and inflammation occurring at the BBB have a key, mainly a deleterious role in the pathophysiology of ischemic stroke. Neddylation is a ubiquitylation-like pathway that is critical in various cellular functions by conjugating neuronal precursor cell-expressed developmentally down-regulated protein 8 (NEDD8) to target proteins. However, the roles of neddylation pathway in ischemic stroke remain elusive. Here, we report that NEDD8 conjugation increased during acute phase after ischemic stroke and was present in intravascular and intraparenchymal neutrophils. Inhibition of neddylation by MLN4924, also known as pevonedistat, inactivated cullin-RING E3 ligase (CRL), and reduced brain infarction and improved functional outcomes. MLN4924 treatment induced the accumulation of the CRL substrate neurofibromatosis 1 (NF1). By using virus-mediated NF1 silencing, we show that NF1 knockdown abolished MLN4924-dependent inhibition of neutrophil trafficking. These effects were mediated through activation of endothelial P-selectin and intercellular adhesion molecule-1 (ICAM-1), and blocking antibodies against P-selectin or anti–ICAM-1 antibodies reversed NF1 silencing-induced increase in neutrophil infiltration in MLN4924-treated mice. Furthermore, we found that NF1 silencing blocked MLN4924-afforded BBB protection and neuroprotection through activation of protein kinase C δ (PKCδ), myristoylated alanine-rich C-kinase substrate (MARCKS), and myosin light chain (MLC) in cerebral microvessels after ischemic stroke, and treatment of mice with the PKCδ inhibitor rottlerin reduced this increased BBB permeability. Our study demonstrated that increased neddylation promoted neutrophil trafficking and thus exacerbated injury of the BBB and stroke outcomes. We suggest that the neddylation inhibition may be beneficial in ischemic stroke.
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影响因子：
15.9
作者：
Lasater, Elisabeth A.;Li, Fang;Ingram, David A., Jr.
通讯作者：
Ingram, David A., Jr.
影响因子：
16
作者：
Brownell, James E.;Sintchak, Michael D.;Dick, Lawrence R.
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
Nam, Do-Hyun
影响因子：
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作者：
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Allan, Stuart M.