Microglial TREM2 Interactome in Alzheimers Disease

阿尔茨海默病中的小胶质细胞 TREM2 相互作用组

• 批准号: 9982605
• 负责人: TAE-WAN KIM
• 金额: $ 52.46万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9982605
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid beta-Protein; Animal Model; Biological; Biological Process; Biology; Brain; Cell Line; Cells; Code; Complex; Disease; Enzymes; Etiology; Genetic Variation; Human; Immune; Knowledge; Label; Ligands; Mass Spectrum Analysis; Mediating; Methods; Microglia; Modeling; Molecular; Pattern; Peroxidases; Play; Protein-Protein Interaction Map; Proteins; Proteomics; Regulation; Research; Research Priority; Risk; Role; Signal Transduction; TREM2 gene; Techniques; Testing; Variant; Work; amyloid pathology; ascorbate; base; commune; induced pluripotent stem cell; knock-down; novel; protein complex; protein protein interaction; receptor; response; risk variant; three-dimensional modeling

Triggering receptor expressed on myeloid cells 2 (TREM2) is an immune modulatory receptor expressed in microglial cells in the brain. Coding variants in TREM2 have been identified as risk factors for Alzheimer's disease (AD). Previous studies have extensively demonstrated that TREM2 plays a central role in microglia activation/survival and amyloid pathology in various cell-based and animal models of AD. Despite evidence demonstrating the functional importance of TREM2 in microglial biology relevant to AD, we currently do not have a complete mechanistic understanding of microglial TREM2 signaling, due in part to the lack of a comprehensive knowledge of TREM2-bearing molecular complex(es) in microglial cells. Most of the focus has been on the interaction of TREM2 with DNAX-activating protein 12 (DAP12), which appears to serve as a communal platform for downstream signaling. Given the complexity and high disease relevance of TREM2-associated biological processes, it is conceivable that additional molecular components may interact with TREM2 and contribute substantially to the regulation of relevant microglial functions. Therefore, we are proposing an unbiased proteomic approach to identify novel components of TREM2-harboring protein complex(es), not yet implicated in TREM2 signaling. Moreover, certain protein-protein-interactions involving TREM2 may be regulated in response to TREM2 receptor stimulation and/or affected by AD-associated TREM2 risk variants. Since conventional protein complex isolation methods are disruptive and often severely affect the stability of the complex, our experimental approach will use an enzyme-catalyzed "proximity labeling" technique to investigate the protein interactions in living cells. Specifically, the ascorbate peroxidase (APEX2)-based proximity-tagging method combined with mass spectrometry will identify proximal endogenously interacting proteins of TREM2 in the intact microglial cell. Successful completion of the proposed research will fill a critical gap in our understanding of the complex biological regulation of TREM2. Our proposal will establish a protein-protein interaction map of TREM2 in microglial cells and may discover novel TREM2-interacting proteins that are critical for TREM2-mediated microglial functions relevant to AD.

骨髓细胞表达的触发受体 2 (TREM2) 是一种免疫调节受体，表达于 大脑中的小胶质细胞。 TREM2 的编码变异已被确定为阿尔茨海默病的危险因素 疾病（AD）。先前的研究已广泛证明 TREM2 在小胶质细胞中发挥着核心作用 各种基于细胞和动物的 AD 模型中的激活/存活和淀粉样蛋白病理学。尽管有证据 证明 TREM2 在与 AD 相关的小胶质细胞生物学中的功能重要性，我们目前还没有 对小胶质细胞 TREM2 信号传导的完整机制了解，部分原因是缺乏全面的 小胶质细胞中携带 TREM2 的分子复合物的知识。大部分的焦点都集中在 TREM2 与 DNAX 激活蛋白 12 (DAP12) 的相互作用，后者似乎充当公共平台 用于下游信令。鉴于 TREM2 相关生物的复杂性和高度疾病相关性 过程中，可以想象额外的分子成分可能与 TREM2 相互作用并有助于 显着影响相关小胶质细胞功能的调节。因此，我们提出一个公正的 蛋白质组学方法鉴定 TREM2 携带蛋白复合物的新成分，尚未涉及 TREM2 信号传导。此外，某些涉及 TREM2 的蛋白质间相互作用可能会受到调节 TREM2 受体刺激和/或受 AD 相关 TREM2 风险变异影响。由于常规 蛋白质复合物分离方法具有破坏性，常常严重影响复合物的稳定性，我们的 实验方法将使用酶催化的“邻近标记”技术来研究蛋白质 活细胞中的相互作用。具体来说，基于抗坏血酸过氧化物酶（APEX2）的邻近标记方法 结合质谱分析将鉴定完整的 TREM2 近端内源性相互作用蛋白 小胶质细胞。成功完成拟议的研究将填补我们理解的一个关键空白 TREM2 的复杂生物调节。我们的提案将建立 TREM2 的蛋白质-蛋白质相互作用图 在小胶质细胞中，可能会发现新的 TREM2 相互作用蛋白，这些蛋白对于 TREM2 介导至关重要 与 AD 相关的小胶质细胞功能。