The COP9 signalosome reduces neuroinflammation and attenuates ischemic neuronal stress in organotypic brain slice culture model

In the context of studying the role of the COP9 signalosome (CSN) in neuroinflammation and ischemic neuronal damage, we studied the effect of the cullin NEDDylation state-modifying drugs MLN4924 and CSN5i-3 in BV2 microglial cells, an immortalized murine cell line featuring many of the characteristics of primary microglia. Owing to its potent inhibitory effect on the NEDDylation cascade, MLN4924 exhibits a CSN5-like anti-inflammatory activity. Csn5i-3 is a small molecule inhibitor that specifically binds to CSN5, while it resides in the CSN holocomplex and blocks its deNEDDylase activity, thus leading to an accumulation of NEDDylated cullins. We performed untargeted mass spectrometry-based proteomics of the cell lysates after treating BV2 cells with these drugs under basal culture stress conditions. The proteomic analysis revealed that MLN4924 and CSN5i-3 substantially altered the microglial proteome.

在本研究中，我们探讨了大分子复合体COP9信号体（CSN）在神经炎症和缺血性神经元损伤中的作用。针对调控Cullin NEDDyl化状态的药物MLN4924和CSN5i-3对BV2小鼠小胶质细胞的影响进行研究，该细胞系为永生化细胞，具有初级小胶质细胞的多项特征。鉴于MLN4924对NEDDyl化级联反应的强大抑制作用，其表现出类似于CSN5的抗炎活性。CSN5i-3是一种小分子抑制剂，能特异性结合CSN5，并在CSN全复合体中定位，从而阻断其去NEDDyl化活性，导致NEDDyl化Cullin的积累。我们在基础培养压力条件下，使用这些药物处理BV2细胞后，对其细胞裂解物进行了非靶向质谱蛋白质组学分析。蛋白质组学分析揭示了MLN4924和CSN5i-3显著改变了小胶质细胞的蛋白质组。