Characterization of the structural and molecular properties of propagating tau in vitro and in vivo

体外和体内增殖 tau 的结构和分子特性表征

基本信息

批准号：
10677817
负责人：
Grace Isabella Hallinan
金额：
$ 14.02万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
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：
2022-08-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10677817
关键词：
Acetylation Affect Alzheimer&apos s Disease Amino Acid Sequence Amyloid beta-Protein Biochemical Biological Models Brain Brain Diseases Cell Culture Techniques Cells Central Nervous System Clinical Trials Cryoelectron Microscopy Custom Data Deacetylation Deposition Development Devices Diagnostic Diagnostic Procedure Disease Disease model Exclusion Filament Future Gene Expression Genes Gerstmann-Straussler-Scheinker Disease Goals Human In Vitro Investigation Knowledge Maps Mass Spectrum Analysis Microfluidic Microchips Microfluidics Modeling Molecular Mus Names Nerve Degeneration Neurodegenerative Disorders Neurons Pathogenicity Pathologic Pathology Pattern Phenotype Pick Disease of the Brain Polymorph Post-Translational Protein Processing Property Protein Isoforms Rodent Model Role Specificity Structure System Tauopathies Techniques Testing Therapeutic Training Transgenic Mice Transgenic Organisms Translations Ubiquitination aggregation pathway brain cell conformer corticobasal degeneration design in vitro Model in vivo in vivo Model interest microscopic imaging neurodegenerative dementia neurodegenerative phenotype neuropathology new technology novel novel diagnostics novel therapeutics paired helical filament prevent prion-like protein expression tau Proteins tau aggregation tau mutation tau-1 technology platform transcriptome sequencing

项目摘要

Project summary/abstract. Tau protein accumulates into pathogenic deposits in many neurodegenerative diseases, including Alzheimer disease (AD) and some forms of Gerstmann-Sträussler-Scheinker disease (GSS). Using cryo-electron microscopy (cryo-EM) recent groundbreaking studies have determined the existence of multiple tau conformers, which may differ between tau diseases. The tau aggregates in AD and GSS contain all 6 isoforms of tau, and both have been ultrastructurally determined to contain paired helical filaments (PHFs). However in AD, there exists an ultrastructural polymorph to PHFs, comprised of the same amino acid sequence but a differing interface, named straight filaments (SFs). These SFs are not found in GSS, nor is it known why tau would fold into PHFs versus SFs in AD. This means that therapeutics designed to target specific tau conformers may not be suitable for all tau polymorphs within a disease. Numerous studies have demonstrated that tau acts in a ‘prion-like’ manner, templating the misfolding from pathogenic ‘donor’ to naïve ‘receiver’ tau, in both in vitro and in vivo models of tau diseases. However, how donor tau acts as a template to receiver tau is not yet known; the mechanism(s) of which would provide a myriad of targets to reduce tau propagation and thus disease dissemination throughout the brain. Our study aims to answer the following questions: is the structure of tau conferred from donor to receiver tau in vivo, and therefore are our in vivo models of neurodegeneration valid for investigating tau propagation (aim 1)? How are the numerous post-translational modifications (PTMs) on tau relevant to tau polymorph formation (aim 2b), and are these PTMs recapitulated from donor to receiver tau both in vivo (aim 2a) and in vitro (aim 3)? What are the gene changes that are occurring in a circuit of diseased brain cells, and are these changes spreading to both anterogradely and retrogradely connected cells (aim 3)? Using novel in vivo and in vitro systems including custom microfluidic cell culture devices, cryo-EM imaging and analysis, mass spectrometry and RNAseq, we aim to answer these vital questions and thus identify future targets for preventing tau propagation.

项目摘要/摘要。 Tau 蛋白在许多神经退行性疾病（包括阿尔茨海默病）中积累成致病沉积物使用冷冻电子治疗疾病 (AD) 和某些形式的 Gerstmann-Sträussler-Scheinker 病 (GSS)。显微镜（冷冻电镜）最近的突破性研究已经确定了多个 tau 蛋白的存在 AD 和 GSS 中的 tau 聚集体包含所有 6 种异构体。 tau 蛋白，并且两者都已被超微结构确定含有成对螺旋丝 (PHF)。 AD，存在 PHF 的超微结构多态性，由相同的氨基酸序列组成，但不同的界面，称为直丝（SF），这些 SF 在 GSS 中没有发现，也不知道为什么 tau。 AD 中的 PHF 与 SF 会折叠在一起，这意味着针对特定 tau 蛋白的治疗方法。大量研究表明，顺应体可能并不适合疾病中的所有 tau 多晶型物。 tau 以“类似朊病毒”的方式发挥作用，模拟从致病性“供体”tau 到天真的“接收者”tau 的错误折叠，然而，在 tau 疾病的体外和体内模型中，供体 tau 如何充当受体 tau 的模板。尚不清楚；其机制将提供无数的目标来减少 tau 传播和因此，疾病在整个大脑中传播。我们的研究旨在回答以下问题： tau 的结构是在体内由供体 tau 赋予受体的，因此是我们的体内模型神经退行性变对于研究 tau 传播（目标 1）有效吗？与 tau 多晶型物形成（目标 2b）相关的 tau 修饰（PTM），这些 PTM 是否已概括从供体到受体的 tau 在体内（目标 2a）和体外（目标 3）有哪些基因变化？发生在患病脑细胞的回路中，这些变化是否会扩散到顺行和逆行连接细胞（目标 3）？使用新型体内和体外系统，包括定制微流体细胞培养设备、冷冻电镜成像和分析、质谱分析和 RNAseq，我们的目标是回答这些重要问题问题，从而确定防止 tau 传播的未来目标。