Deep proteome profiling reveals novel pathways associated with proinflammatory and alcohol-induced microglial activation phenotypes
Deep proteome profiling reveals novel pathways associated with proinflammatory and alcohol-induced microglial activation phenotypes
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DOI：
10.1016/j.jprot.2020.103753
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发表时间：
2020-05-30
影响因子：
3.3
通讯作者：
Stevens, Stanley M., Jr.
中科院分区：
文献类型：
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作者：
Guergues, Jennifer;Wohlfahrt, Jessica;Stevens, Stanley M., Jr.
小胶质细胞是大脑的常驻免疫细胞，可以表现出广泛的活化表型，其中许多与中枢神经系统的几种疾病和病症有关，包括与酒精滥用有关的疾病和病症。鉴于定义小胶质细胞活化的全球范围分子变化的复杂性，在酒精暴露的背景下仍然缺乏准确的表型分类。我们采用了一种优化的方法对初级小胶质细胞进行深度定量蛋白质组分析，以表征它们对急性暴露于酒精（乙醇）以及促炎驱动剂和TLR4激动剂LPS的反应。从该分析中，鉴定了5，062种总蛋白质，其中在乙醇和LPS处理组中分别有4，857和4，928种蛋白质可通过无标记定量来定量。这项研究强调了乙醇处理的小胶质细胞中发生的微妙但显著的蛋白质组学变化，这些变化与TLR4激活诱导的强大促炎表型不一致。具体而言，我们的结果表明与炎症相关的几种上游调节剂的抑制，与TLR4介导的促炎表型相比对途径如吞噬作用的相反作用，以及与OXPHOS和脂质稳态相关的蛋白质表达增加相关的潜在代谢转变。数据可通过ProteomeXchange获得，标识符为PXD 14466。意义：酒精滥用对中枢神经系统有显著影响，其中包括神经胶质细胞激活引起的病理生理机制。特别是小胶质细胞，是大脑的常驻免疫细胞，并表现出广泛的活化表型。驱动小胶质细胞活化表型的分子变化是复杂的，尚未在酒精暴露的背景下充分表征。我们的研究突出了迄今为止初级小胶质细胞中酒精诱导的蛋白质组变化的第一个也是最全面的特征，并揭示了由于酒精暴露而改变的新型免疫相关和代谢途径。这项研究的结果为未来的工作提供了重要的基础，旨在了解酒精诱导的小胶质细胞激活的复杂性在体内和其他翻译模型的急性和慢性酒精暴露。
Microglia, the resident immune cells of the brain, can exhibit a broad range of activation phenotypes, many of which have been implicated in several diseases and disorders of the central nervous system including those related to alcohol abuse. Given the complexity of global-scale molecular changes that define microglial activation, accurate phenotypic classification in the context of alcohol exposure is still lacking. We employed an optimized method for deep, quantitative proteome profiling of primary microglia in order to characterize their response to acute exposure to alcohol (ethanol) as well as the pro-inflammatory driver and TLR4 agonist, LPS. From this analysis, 5,062 total proteins were identified where 4,857 and 4,928 of those proteins were quantifiable by label-free quantitation in ethanol and LPS treatment groups, respectively. This study highlights the subtle, yet significant proteomic changes that occur in ethanol-treated microglia, which do not align with the robust pro-inflammatory phenotype induced by TLR4 activation. Specifically, our results indicate inhibition of several upstream regulators associated with inflammation, opposing effects on pathways such as phagocytosis upon comparison to TLR4-mediated pro-inflammatory phenotype, and a potential metabolic shift associated with increased expression of proteins related to OXPHOS and lipid homeostasis. Data are available via ProteomeXchange with identifier PXD14466.Significance: Alcohol abuse has a significant impact on the central nervous system, which includes the pathophysiological mechanisms resulting from glial cell activation. Microglia, in particular, are the resident immune cells of the brain and exhibit a broad range of activation phenotypes. The molecular changes that drive microglial activation phenotype are complex and have yet to be fully characterized in the context of alcohol exposure. Our study highlights the first and most comprehensive characterization of alcohol-induced proteomic changes in primary microglia to date and has shed light on novel immune-related and metabolic pathways that are altered due to alcohol exposure. The results from this study provide an important foundation for future work aimed to understand the complexity of alcohol-induced microglial activation in vivo and other translational models of acute and chronic alcohol exposure.