APEX2-mediated Identification of factors involved in seeding by tau protein

Tau病理以普恩病毒的方式传播被认为是进行性疾病的基础。 Tau聚集物在tauopathy中的积累。在实验系统中，很容易观察到 在转基因动物中，tau聚集体从细胞外到细胞内以及在脑网络内的传播 老鼠。然而，目前尚不清楚这种现象是否是进行性神经变性的原因。 在人类的变态疗法中。我们不知道tau聚集进入细胞的分子机制。 可以复制独特的结构。对相关因素的识别将有助于检验普里恩假说。 进行性神经退行性变。此外，了解这些机制可能有助于 开发新的治疗方法。我们建议使用一种新开发的工具来识别和表征因素 参与tau聚集的基因。抗坏血酸过氧化物酶(APEX2)依赖的邻近标记是一种 细胞内蛋白质的时间分辨生物素化的酶方法。我们将鉴定那些具有 基于亲和纯化和质谱学的标记。这将使我们能够近距离地确定因素 聚集过程开始时靠近tau，在巨噬细胞吞噬或直接将聚集体输送到 牢房内部。我们将使用标准遗传学来跟进命中结果。这项工作可能会对我们的 了解细胞内淀粉样蛋白的传播，并可能形成新的治疗方法的基础 战略。

Propagation of tau pathology in the manner of prions has been hypothesized to underlie the progressive accumulation of tau aggregates in tauopathies. In experimental systems it has been easy to observe propagation of tau aggregation from the outside to the inside of the cell, and within brain networks in transgenic mice. However, it remains unknown whether this phenomenon is the cause of progressive neurodegeneration in human tauopathies. We do not know the molecular mechanisms by which tau aggregates taken into a cell can replicate unique structures. The identification of factors that are involved will help test the prion hypothesis of progressive neurodegeneration. Furthermore, understanding these mechanisms could help with the development of new therapies. We propose to use a newly developed tool to identify and characterize factors that are involved in tau aggregation. Ascorbate peroxidase (APEX2)-dependent proximity labeling is an enzymatic method for time-resolved biotinylation of proteins within a cell. We will identify peptides that have been labeled based on affinity purification and mass spectrometry. This will let us identify factors in close proximity to tau as the aggregation process begins, after macropinocytosis or direct delivery of aggregates to the cell interior. We will use standard genetics to follow up hits. This work could have strong impact on our understanding of the propagation of intracellular amyloids, and could form the basis for new therapeutic strategies.