Deciphering the Molecular Features Underlying LRP1-Mediated Tau Spread

破译 LRP1 介导的 Tau 扩散的分子特征

基本信息

批准号：
10622556
负责人：
Jennifer Nicole Rauch
金额：
$ 38.87万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10622556
关键词：
3-Dimensional Acetylation Affinity Alternative Splicing Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease pathology Alzheimer&apos s disease therapeutic Alzheimer&apos s disease therapy Amber Biochemical Biological Biological Assay Brain Cell Culture Techniques Cells Charge Codon Nucleotides Complex Cryoelectron Microscopy Deposition Development Disease Disease Progression Dissection Electrostatics Elements Evaluation Fluorescence Resonance Energy Transfer Genetic Goals Human In Vitro Induced pluripotent stem cell derived neurons Intervention Knowledge LDL-Receptor Related Protein 1 Learning Ligands Link Lipoprotein Receptor Lysine Maps Mass Spectrum Analysis Measures Mediating Methods Modeling Modification Molecular Mus Neurodegenerative Disorders Pathogenicity Pathologic Pathway interactions Phosphorylation Point Mutation Post-Translational Protein Processing Process Protein Isoforms Proteins Protocols documentation Publishing Regulation Sampling Site Structure Surface Tauopathies Technology Testing Ubiquitin Ubiquitination Work comparison control density effective therapy human disease in vivo innovation insight knock-down novel novel therapeutics prevent protein aggregation protein protein interaction receptor tau Proteins tau aggregation tau interaction tau mutation tau-1 therapeutic target treatment strategy unnatural amino acids unpublished works uptake

项目摘要

Several neurodegenerative diseases, such as Alzheimer’s disease (AD), are characterized by the spread and aggregation of the protein tau. Recently, we identified a cellular receptor, LRP1 (Low-density lipoprotein Receptor-related Protein 1), that regulates the tau spread pathway. Knockdown of LRP1 prevents tau spread in human iPS neurons and the mouse brain, suggesting that the tau-LRP1 interaction could be an important entry point for disease intervention. Unfortunately, a detailed understanding of the tau-LRP1 molecular complex is still lacking. Therefore, the main objective of this project is to define the tau-LRP1 structural interface and discern how post-translation modifications (PTMs) to tau’s structure influence tau uptake and spread. In preliminary work, we have developed protocols to purify and measure interactions between tau and LRP1. We have established cellular platforms to model tau propagation and have shown that this process can be influenced by tau PTMs. To fully develop this work, we propose three aims. In Aim 1, we will use TR-FRET to establish in vitro affinities between tau and LRP1 and mass spectrometry to map the protein-protein interface. In Aim 2, we will look at how tau phosphorylation can influence the tau-LRP1 complex and what effect this has on tau spread and aggregation. In Aim 3, we will focus on tau PTMs that alter lysine residues. We will assess if ubiquitination or acetylation can impact the tau-LRP1 interaction, if they influence tau aggregation, and if they promote or inhibit tau uptake and seeding in cells. The innovative experimental methods and comprehensive analyses outlined herein will provide important mechanistic insight and develop our understanding of pathogenic tau regulation in AD. This will be an essential first step forward for the development and evaluation of potential AD therapeutics.

几种神经退行性疾病，例如阿尔茨海默氏病（AD），其特征是扩散和蛋白质的聚集。最近，我们鉴定了一个细胞受体LRP1（低密度脂蛋白与受体相关的蛋白质1），可调节tau扩散途径。 LRP1的敲低可防止Tau传播在人IPS神经元和小鼠大脑中，表明tau-lrp1相互作用可能很重要疾病干预的入口点。不幸的是，对tau-lrp1分子复合物的详细理解仍然缺乏。因此，该项目的主要目的是定义tau-lrp1结构界面和辨别翻译后修改（PTM）如何影响tau的结构影响tau的吸收和扩散。在初步工作，我们开发了净化和衡量tau和LRP1之间的相互作用的协议。我们已经建立了蜂窝平台来建模tau的传播，并表明该过程可能受到影响由tau ptms。为了充分开发这项工作，我们提出了三个目标。在AIM 1中，我们将使用Tr-Fret在 Tau和LRP1和质谱之间的体外亲和力以绘制蛋白质蛋白质界面。在AIM 2中，我们将研究tau磷酸化如何影响tau-lrp1复合物，以及这对tau蔓延的影响和聚合。在AIM 3中，我们将专注于改变赖氨酸保留的tau PTM。我们将评估泛素化是否或乙酰化会影响tau-lrp1的相互作用，如果它们影响tau聚集，如果它们促进或抑制 tau摄取并在细胞中播种。概述的创新实验方法和全面分析以下将提供重要的机械洞察力，并发展我们对致病性TAU调节的理解广告。这将是对潜在AD疗法开发和评估的重要第一步。