The NEDD8-activating enzyme inhibitor MLN4924 reduces ischemic brain injury in mice.
Huilin YuHaiyu LuoLuping ChangShisheng WangXue GengLijing KangYi ZhongYongliang CaoRanran WangXing YangYuanbo ZhuMei-Juan ShiYue HuZhongwang LiuXuhui YinYun Wei RanHao YangWenying FanBing-Qiao ZhaoPublished in: Proceedings of the National Academy of Sciences of the United States of America (2022)
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.
Keyphrases
- blood brain barrier
- cerebral ischemia
- signaling pathway
- brain injury
- lps induced
- atrial fibrillation
- oxidative stress
- protein kinase
- pi k akt
- subarachnoid hemorrhage
- nuclear factor
- diabetic rats
- high fat diet induced
- high glucose
- endothelial cells
- inflammatory response
- type diabetes
- binding protein
- staphylococcus aureus
- amino acid
- functional connectivity
- resting state
- mesenchymal stem cells
- multiple sclerosis
- coronary artery
- stem cells
- drug induced
- cell therapy
- cell proliferation
- biofilm formation
- small molecule
- tyrosine kinase
- ischemia reperfusion injury
- weight loss
- newly diagnosed