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：
10613553
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/10613553
关键词：
Acceleration 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 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 Small Interfering RNA Specimen Symptoms Tauopathies Testing Therapeutic Transfection Treatment Efficacy Vesicle abnormally phosphorylated tau aged antagonist brain tissue candidate identification case control chronic traumatic encephalopathy comparison control corticobasal degeneration differential expression exosome extracellular vesicles fortification gray matter in vitro Assay in vitro activity in vivo induced pluripotent stem cell microvesicles mind control mouse model multi-electrode arrays multidisciplinary non-demented protein protein interaction recruit sensor sex tau Proteins tau aggregation 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，由过度磷酸化微管相关的细胞内聚集体组成 tau 蛋白 (tau) 是迄今为止与阿尔茨海默病 (AD) 临床症状最相关的病理学。新的证据表明细胞外囊泡（EV，例如外泌体和微泡），转移病理性tau蛋白在细胞间作为载体，传播tau蛋白病理学。当务之急是找到脑源性 EV 的分子基础，可能关键调节神经元对 EV 的摄取和聚集 EV 和/或受体神经元中的 tau 蛋白。我们最近建立了将电动汽车与电动汽车隔离的方法人类大脑样本并成功进行了蛋白质组分析。我们发现选择性分子来自 EV 蛋白质组学数据集的研究人员能够将人类 AD-EV 与健康对照 (CTRL)-EV 区分开来机器学习分析的准确度为 88%，确认了 AD-EV 分子的致病特征。此外，我们令人兴奋的初步数据表明 AD-EV 具有显着更高的 tau 播种通过 FRET 传感器 tau 播种测定与 CTRL-EV 的活性进行比较，其中 EV 分子子集显示与 tau 播种活性显着相关。该拟议项目将巩固这些初步成果发现 tau 蛋白病之间介导 tau 聚集及其播种的趋同或特定机制通过脑源性 EV 样本的蛋白质组学和生物检查来摄取 EV。为了满足这个面对挑战，我们组建了一个多学科的研究团队，他们在在生物学（Ikezu）、蛋白质组学（Emili）以及生物工程和生物信息分析（Issador）方面取得的成就。在 Aim1 中，我们将检查来自 240 个新大脑样本的 EV 样本（40 AD、40 CTE、40 LBD、40 PSP、40 CBD 和 40 CTRL) 用于基于质谱的精密蛋白质组学和 tau 相互作用组学，并进行分析通过机器学习方法获得这些数据集。目标 2 将使用以下方法检查 tau 传播的效率从 5 种不同 tau 病和对照病例的相同供体中分离出 tau 原纤维、寡聚体和 EV 使用基于 FRET 的 tau 接种测定和原代培养小鼠皮质的 EV 摄取进行体外和体内实验体外的神经元。 EV 相关 tau 蛋白将通过基于生化和显微镜的进一步表征分析它们的构象和翻译后变化。我们将评估 tau 的差异使用我们最近的研究，将来自不同 tau 病大脑的 tau 种子颅内注射后的传播建立小鼠模型。 Aim3 将识别最有可能参与 EV 摄取和 tau 蛋白的候选分子通过蛋白质组数据集（目标 1）和生物数据集（目标 2）的生物信息学分析进行播种活动。我们然后将测试已识别分子对 EV 摄取、tau 播种活性和神经元的功能作用体外发射活动。候选分子将被基因沉默或拮抗剂特异性靶向因其在体外和体内阻止 tau 增殖的治疗潜力。成功识别责任人 tau 增殖分子将成为了解 EV 介导的疾病进展的基础。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

ATP1A3 as a target for isolating neuron-specific extracellular vesicles from human brain and biofluids.

ATP1A3 作为从人脑和生物体液中分离神经元特异性细胞外囊泡的靶标。

DOI：
发表时间：
2023-09-15
期刊：
影响因子：
13.6
作者：
You, Yang;Zhang, Zhengrong;Sultana, Nadia;Ericsson, Maria;Martens, Yuka A;Sun, Min;Kanekiyo, Takahisa;Ikezu, Seiko;Shaffer, Scott A;Ikezu, Tsuneya
通讯作者：
Ikezu, Tsuneya

Enrichment of Phosphorylated Tau (Thr181) and Functionally Interacting Molecules in Chronic Traumatic Encephalopathy Brain-derived Extracellular Vesicles.

慢性创伤性脑病脑源性细胞外囊泡中磷酸化 Tau (Thr181) 和功能相互作用分子的富集。

DOI：
发表时间：
2021-09
期刊：
影响因子：
0
作者：
Muraoka, Satoshi;Lin, Weiwei;Takamatsu;Hu, Jianqiao;Ikezu, Seiko;DeTure, Michael A;Dickson, Dennis W;Emili, Andrew;Ikezu, Tsuneya
通讯作者：
Ikezu, Tsuneya

Extracellular vesicles as personalized medicine.

细胞外囊泡作为个性化医疗。

DOI：
发表时间：
2023-06
期刊：
影响因子：
10.6
作者：
Beetler, Danielle J;Di Florio, Damian N;Bruno, Katelyn A;Ikezu, Tsuneya;March, Keith L;Cooper Jr, Leslie T;Wolfram, Joy;Fairweather, DeLisa
通讯作者：
Fairweather, DeLisa

Antisense oligonucleotide-based targeting of Tau-tubulin kinase 1 prevents hippocampal accumulation of phosphorylated tau in PS19 tauopathy mice.

基于反义寡核苷酸的 Tau 微管蛋白激酶 1 靶向可防止 PS19 tau 蛋白病小鼠海马中磷酸化 tau 蛋白的积累。

DOI：
发表时间：
2023-10-19
期刊：
影响因子：
7.1
作者：
Yukawa, Kayo;Yamamoto;Cafaro, Louis;Hong, Christine;Kamme, Fredrik;Ikezu, Tsuneya;Ikezu, Seiko
通讯作者：
Ikezu, Seiko

Recommendations for reproducibility of cerebrospinal fluid extracellular vesicle studies.

关于脑脊液细胞外囊泡研究重现性的建议。

DOI：
发表时间：
2024-01
期刊：
影响因子：
0
作者：
Sandau, Ursula S;Magaña, Setty M;Costa, Júlia;Nolan, John P;Ikezu, Tsuneya;Vella, Laura J;Jackson, Hannah K;Moreira, Lissette Retana;Palacio, Paola Loreto;Hill, Andrew F;Quinn, Joseph F;Van Keuren;McFarland, Trevor J;Palade
通讯作者：
Palade