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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：
10834533
负责人：
Jennifer Nicole Rauch
金额：
$ 5.09万
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/10834533
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Proteins tau aggregation tau interaction tau mutation tau-1 therapeutic target treatment strategy unnatural amino acids unpublished works uptake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.

一些神经退行性疾病，例如阿尔茨海默病 (AD)，其特点是传播和传播 tau 蛋白的聚集。最近，我们鉴定了一种细胞受体，LRP1（低密度脂蛋白）受体相关蛋白 1)，调节 tau 扩散途径。敲低 LRP1 可防止 tau 扩散在人类 iPS 神经元和小鼠大脑中，表明 tau-LRP1 相互作用可能是一个重要的疾病干预的切入点。不幸的是，对 tau-LRP1 分子复合物的详细了解仍然缺乏。因此，本项目的主要目标是定义tau-LRP1结构界面并了解 tau 结构的翻译后修饰 (PTM) 如何影响 tau 的摄取和传播。在在前期工作中，我们开发了纯化和测量 tau 和 LRP1 之间相互作用的方案。我们已经建立了细胞平台来模拟 tau 传播，并表明该过程可以受到影响通过 tau PTM。为了充分开展这项工作，我们提出三个目标。在目标 1 中，我们将使用 TR-FRET 建立 tau 和 LRP1 之间的体外亲和力以及质谱法来绘制蛋白质-蛋白质界面。在目标 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 扩散有何影响和聚合。在目标 3 中，我们将重点关注改变赖氨酸残基的 tau PTM。我们将评估是否泛素化或乙酰化可以影响 tau-LRP1 相互作用，如果它们影响 tau 聚集，并且如果它们促进或抑制 tau 蛋白在细胞中的摄取和播种。概述的创新实验方法和综合分析本文将提供重要的机制见解并加深我们对致病性 tau 调节的理解广告。这将是开发和评估潜在 AD 疗法的重要的第一步。