Novel Proteomic Approaches for the Study of Alcohol Neuropathology

 描述（由申请方提供）：过量乙醇消耗诱导神经毒性，可能涉及神经炎症。小胶质细胞是中枢神经系统（CNS）中的主要免疫细胞，并且被认为是神经炎症相关神经毒性的主要贡献者;然而，小胶质细胞在酒精诱导的神经元功能障碍中的作用尚未完全理解，特别是在神经元-小胶质细胞相互作用的背景下。在这项提议中，我们将采用新的蛋白质组学方法来识别和量化差异蛋白质表达，以测试我们的工作假设，即小胶质细胞与周围神经元的相互作用在酒精诱导的小胶质细胞活化和随后的神经毒性中起着关键作用。我们建议使用一种新的Super-SILAC方法在慢性乙醇暴露的体内模型中确定乙醇诱导的小胶质细胞反应。通过Super- SILAC蛋白质组学分析鉴定的变化将映射到miRNA差异表达，以建立以非编码RNA及其在蛋白质表达中的调节作用为特征的全球规模的机制网络。我们提出的研究结果将进一步深入了解小胶质细胞在乙醇诱导的神经变性中的作用，并确定新的治疗策略的潜在靶点，用于治疗或预防由过度和长期饮酒引起的神经元损伤。

 DESCRIPTION (provided by applicant): Excessive ethanol consumption induces neurotoxicity that may involve neuroinflammation. Microglia are the primary immune cells in the central nervous system (CNS) and have been implicated as a major contributor to neuroinflammation-related neurotoxicity; however, the role of microglia in alcohol- induced neuronal dysfunction is not fully understood especially in the context of neuron-microglia interplay. In this proposal, we will employ novel proteomic methods to identify and quantify differential protein expression to test our working hypothesis that the interplay of microglia with surrounding neurons plays a pivotal role in alcohol-induced microglial activation and subsequent neurotoxicity. We propose to determine the ethanol-induced microglial response in an in vivo model of chronic ethanol exposure using a novel Super-SILAC approach. Changes identified by Super- SILAC proteomic profiling will be mapped to miRNA differential expression to establish a global-scale mechanistic network featuring non-coding RNAs and their regulatory role in protein expression. The results from our proposed studies will provide further insight into the role of microglia in ethanol- induced neurodegeneration and identify potential targets for new therapeutic strategies for the treatment or prevention of neuronal injury brought about by excessive and long-term alcohol consumption.