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Molecular characterization of extracellular vesicles for the spread of misfolded tau protein

错误折叠 tau 蛋白扩散的细胞外囊泡的分子特征

基本信息

批准号：
10404919
负责人：
Tsuneya Ikezu
金额：
$ 61.14万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-15 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10404919
关键词：
Age Alzheimer&apos s Disease Alzheimer&apos s disease brain Atomic Force Microscopy Binding Biochemical Bioinformatics Biological Biological Assay Biomedical Engineering Biosensor Brain C57BL/6 Mouse Cells Clinical Pathology Data Data Set Databases Disease Disease Progression Endocytosis Energy Transfer Enzyme-Linked Immunosorbent Assay Evaluation Foundations Gene Silencing Genes Goals Hippocampus (Brain)Human In Vitro Induced pluripotent stem cell derived neurons Inflammation Injections Lewy Body Dementia MAPT gene Machine Learning Mass Spectrum Analysis Mediating Methods Microscopy Molecular Molecular Conformation Molecular Target Mus Neurofibrillary Tangles Neurons Pathogenicity Pathologic Pathology Post-Translational Protein Processing Progressive Supranuclear Palsy Property Proteins Proteome Proteomics Reagent Research Personnel Role Sampling Seeds Small Interfering RNA Specimen Symptoms Tauopathies Testing Therapeutic Transfection Treatment Efficacy Vesicle abnormally phosphorylated tau aged antagonist base brain tissue case control chronic traumatic encephalopathy corticobasal degeneration differential expression disorder control exosome extracellular vesicles gray matter in vitro Assay in vitro activity in vivo induced pluripotent stem cell microvesicles mouse model multi-electrode arrays multidisciplinary non-demented protein protein interaction recruit sensor sex tau Proteins tau aggregation tau phosphorylation tau-1 transmission process uptake

项目摘要

Neurofibrillary tangles, composed of intracellular aggregates of hyperphosphorylated microtubule-associated protein tau (tau), are by far the most correlated pathology for clinical symptoms of Alzheimer disease (AD). Emerging evidence suggests that extracellular vesicles (EVs, such as exosomes and microvesicles), transfer pathological tau between cells as vehicles, propagating tau pathology. It is urgently important to find the molecular basis of brain-derived EVs, which may critically regulate EV uptake by neurons and aggregation of tau protein in EVs and/or recipient neurons. We have recently established the method for isolating EVs from human brain samples and successfully performed their proteomic profiling. We found that selective molecules from the EV proteomics datasets were able to differentiate human AD-EV from healthy control (CTRL)-EV with 88% accuracy by machine learning analysis, confirming pathogenic character of AD-EV molecules. Furthermore, our exciting preliminary data have shown that AD-EV have significantly higher tau seeding activity compared to CTRL-EV by FRET sensor tau seeding assay with subsets of EV molecules showing significant association with tau seeding activity. This proposed project will fortify these preliminary results and find the converging or specific mechanisms among tauopathies for mediating tau aggregation and its seeding via EV uptake through proteomics and biological examination of brain-derived EV samples. To meet this challenge, we assembled a multi-disciplinary team of investigators who have a strong record of accomplishments in biologic (Ikezu), proteomic (Emili) and bioengineering and bioinformatic analysis (Issador). In Aim1, we will examine EV samples from 240 new brain specimens (40 AD, 40 CTE, 40 LBD, 40 PSP, 40 CBD and 40 CTRL) for precision mass-spectrometry-based proteomics and tau-interactomes, and analyze those datasets by the machine learning approach. Aim 2 will examine the efficiency of tau propagation using tau fibrils, oligomers and EVs isolated from the same donors of the 5 different tauopathies and control cases in vitro and in vivo using FRET-based tau seeding assay and EV uptake by primary cultured mouse cortical neurons in vitro. EV-associated tau will be further characterized by the biochemical and microscopy-based analysis for their conformational and posttranslational changes. We will evaluate the difference in tau propagation after the intracranial injection of the tau seeds from different tauopathy brains using our recently established mouse models. Aim3 will identify candidate molecules most likely involved in EV uptake and tau seeding activity by bioinformatic analysis of the proteome dataset (Aim 1) and biological datasets (Aim 2). We will then test the functional roles of the identified molecules on EV uptake, tau seeding activities and neuronal firing activities in vitro. The candidate molecules will be specifically targeted by gene silencing or antagonists for their therapeutic potential to halt tau propagation in vitro and in vivo. Successful identification of responsible molecules for tau propagation will serve as a foundation for understanding EV-mediated disease progression.

神经原纤维缠结，由细胞内过度磷酸化的微管相关聚集体组成