Chemical Tools to Target TREM2 in Alzheimer's Disease

项目摘要/摘要 最近的全基因组关联研究和使用阿尔茨海默病(AD)小鼠模型的研究已经 发现一种小胶质受体TREM2(在髓系细胞上表达的触发受体2)是一种新的治疗方法 AD的目标。为制定有效的治疗策略，全面了解TREM2的发病机制 函数是必需的。然而，由于与TREM2相关的复杂性和缺乏，它一直具有挑战性 研究工具。尽管有许多遗传工具可用，但很少有化学生物学工具被开发出来 研究TREM2生物学。长期的研究目标是了解TREM2的详细机制 在AD发病机制中的作用，并确定新的治疗靶点和策略。的主要目标是 这项建议是开发化学生物学工具来研究TREM2糖基化，鉴定sTREM2的受体， 并探讨了TREM2的激活机制。这一目标将通过三个目标来实现。目标1 是开发一种健壮的合成方法来生产具有均一多聚糖的TREM2胞外结构域，并检测 糖基化对TREM2折叠、稳定性和与其配体结合的影响。目标2是开发sTREM2 带有光反应基团的蛋白质交联探针和亲和纯化的浓缩标签。这些 将应用探针来识别sTREM2受体。目标3是开发受限制的多肽来结合和 激活TREM2信号，从而从机制上理解TREM2的激活。这项提议是创新的。 因为1)通过末端唾液酸化改变糖基化模式可以影响TREM2的创新假说 功能，挑战现有的TREM2糖基化观点和2)它将产生几个新的化学生物学 科学界可用来研究TREM2的工具。这项提议意义重大，因为1)它将增加 我们对复杂的TREM2生物学的理解，2)它将识别一种sTREM2受体，一种潜在的新的 AD的治疗靶点，以及3)它将产生TREM2的限制性多肽激动剂，可以进一步优化 作为临床前试验的候选药物。

PROJECT SUMMARY/ABSTRACT Recent genome-wide association studies and studies using mouse models of Alzheimer’s disease (AD) have identified a microglial receptor TREM2 (triggering receptor expressed on myeloid cells 2) as a new therapeutic target of AD. To develop effective therapeutic strategies, a comprehensive mechanistic understanding of TREM2 functions is needed. However, it has been challenging due to the complexity associated with TREM2 and a lack of research tools. Although many genetic tools are available, few chemical biology tools have been developed to study TREM2 biology. The long-term research goal is to understand the detailed mechanism of TREM2 functions during AD pathogenesis and identify novel therapeutic targets and strategies. The main objective of this proposal is to develop chemical biology tools to study TREM2 glycosylation, identify the receptor of sTREM2, and investigate the activation mechanism of TREM2. This objective will be accomplished with three aims. Aim 1 is to develop a robust synthetic method to produce TREM2 ectodomain with homogenous glycans and examine the effect of glycosylation on TREM2 folding, stability, and binding to its ligands. Aim 2 is to develop sTREM2 probes with photo-reactive groups for protein cross-linking and enrichment tags for affinity purification. These probes will be applied to identify the sTREM2 receptor. Aim 3 is to develop constrained peptides to bind and activate TREM2 signaling, allowing a mechanistic understanding of TREM2 activation. The proposal is innovative because 1) innovative hypothesis that altered glycosylation patterns by terminal sialylation can affect TREM2 functions, challenging the existing view of TREM2 glycosylation and 2) it will yield several novel chemical biology tools available to the scientific community to study TREM2. The proposal is significant because 1) it will increase our understanding of the complex TREM2 biology, 2) it will identify a sTREM2 receptor, a potential new therapeutic target of AD, and 3) it will yield constrained peptide agonists of TREM2 that can be further optimized as drug candidates for preclinical testing.