Human Cerebrospinal Fluid Extracellular Vesicles: Utility as Disease Specific Biomarkers and Impact on Alzheimer's Disease Pathology

人脑脊液细胞外囊泡：作为疾病特异性生物标志物的用途及其对阿尔茨海默病病理学的影响

• 批准号: 10661249
• 负责人: JULIE Anne SAUGSTAD
• 金额: $ 73.77万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661249
• 关键词: Age; Alleles; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Alzheimer’s disease biomarker; American; Amyloid beta-42; Amyloid beta-Protein; Apolipoprotein E; Astrocytes; Autopsy; Biogenesis; Biological Assay; Biological Factors; Biological Markers; Brain; Cause of Death; Cells; Cerebrospinal Fluid; Classification; Cognitive; Data; Dementia; Dependence; Disease; Enzyme-Linked Immunosorbent Assay; Etiology; Female; Flow Cytometry; Functional disorder; Gene Targeting; Genetic; Genotype; Goals; High Prevalence; Hippocampus; Human; Immunohistochemistry; In Vitro; Individual; Information Theory; Inherited; Living Donors; Luciferases; Measurement; Measures; Memory Loss; Messenger RNA; Methods; MicroRNAs; Molecular; Molecular Sieve Chromatography; Neurodegenerative Disorders; Neurofibrillary Tangles; Neuroglia; Neurologic; Neurons; Parkinson Disease; Participant; Pathogenesis; Pathologic Processes; Patients; Persons; Process; Progressive Disease; Proteins; Proteomics; Qualifying; Regulation; Reporter; Research Personnel; Research Priority; Risk; Risk Factors; Surface; Time; Transfection; Transmission Electron Microscopy; Vesicle; Western Blotting; Woman; abnormally phosphorylated tau; apolipoprotein E-4; biomarker identification; brain cell; candidate identification; candidate marker; care costs; cell type; cerebral atrophy; comparison control; cost; disorder control; exosome; extracellular vesicles; frontal lobe; genetic risk factor; high risk; human old age (65+); improved; in vivo; male; men; microRNA biomarkers; microvesicles; mild cognitive impairment; mind control; new therapeutic target; particle; payment; protein biomarkers; protein expression; response; sex; specific biomarkers; statistics; tau Proteins; tau-1

PROJECT SUMMARY Alzheimer's disease (AD) is the most common form of dementia and the fifth leading cause of death for those age 65 and older. Costs of care for individuals with AD are substantial: total payments in 2022 are estimated at $321 billion, and costs will increase by $1 billion each year. AD risk factors include age, sex, and genetics. Of Americans living with AD, 4 million are women and 2.5 million are men, and the apolipoprotein E (APOE) -e4 allele is the most important genetic risk factor for sporadic AD. In addition, female APOE-e4 carriers are more likely to progress from mild cognitive impairment (MCI) to AD, have more brain atrophy and memory loss, and develop AD more frequently than age matched males. Hallmark features of AD include the buildup of amyloid beta (Aβ) and abnormal phosphorylation of Tau, resulting in plaques and neurofibrillary tangles in the brain. Importantly in AD, disruptions to extracellular vesicle (EV) biogenesis result in altered EV miRNA and protein cargo, and neuronal EVs in AD contain and propagate both Aβ and Tau. Our previous studies identified miRNAs in human cerebrospinal fluid (CSF) from living donors that discriminate AD patients from healthy control (CTL) participants, and we found that combining CSF miRNAs plus CSF Aβ42:total Tau measurements improves classification of AD and MCI vs. CTL. In the studies proposed herein, we focus on the utility of CSF EVs and their cargo as more robust biomarkers for AD, that show distinct differences in females vs. males and those with the APOE-e4 vs. APOE-e3 genotype. Our pilot data in human CSF EVs and postmortem brain shows that: i) the size of membranous particles is larger in AD vs. CTL CSF; ii) CSF EVs have surface marker proteins that are unique to AD and/or Parkinson's disease (PD) vs. CTL; iii) CSF EV miRNAs have improved predictive power for AD vs. total CSF; iv) CSF EV miRNAs show sex and APOE dependent changes in expression; v) there is increased miR-16-5p in female AD CSF EVs vs. CTL; and vi) a predicted gene target of miR-16-5p, SNAP-25, is decreased in female AD hippocampus. Together, these studies support that sex and APOE-e4 both contribute to changes in AD CSF EVs, which may account in part for the higher prevalence of AD in females, and the increased risk for AD in people with the APOE-e4 genotype. Thus, we propose to establish the physical features and molecular cargo of AD CSF EVs vs. CTL, within the context of sex and APOE genotype. We will use our Information Theory method to identify the strongest dependencies that can distinguish AD from PD from CTL, and to identify candidate biomarkers for high-risk AD. We include male and female PD to identify CSF EV surface markers and cargo that are unique from AD. We will then identify and verify gene targets of CSF EV miRNAs that are relevant to AD. Together, our studies will establish specific biomarkers for AD by combining the physical features, surface marker profiles, miRNAs, and proteins in CSF EVs. They will also establish the neurologically relevant gene targets of AD EV miRNAs in human brain, which may contribute to the processes underlying AD pathogenesis and serve as novel therapeutic targets for AD.

项目概要 阿尔茨海默病 (AD) 是最常见的痴呆症，也是痴呆症患者的第五大死因 65 岁及以上的 AD 患者的护理费用很高：预计 2022 年的总付款额为 3210 亿美元，AD 风险因素包括年龄、性别和遗传，费用每年将增加 10 亿美元。 患有 AD 的美国人中，400 万是女性，250 万是男性，载脂蛋白 E (APOE) -e4 等位基因是散发性AD最重要的遗传危险因素，此外，女性APOE-e4携带者较多。 可能从轻度认知障碍 (MCI) 发展为 AD，有更多的脑萎缩和记忆丧失，并且 AD 的标志性特征包括淀粉样蛋白的积累。 β (Aβ) 和 Tau 蛋白异常磷酸化，导致大脑中出现斑块和神经原纤维缠结。 重要的是，在 AD 中，细胞外囊泡 (EV) 生物发生的破坏会导致 EV miRNA 和蛋白质发生改变 我们之前的研究发现，AD 中的货物和神经元 EV 含有并传播 Aβ 和 Tau。 活体捐献者脑脊液 (CSF) 中的 miRNA 可区分 AD 患者和健康人 对照 (CTL) 参与者，我们发现将 CSF miRNA 与 CSF Aβ42 相结合：总 Tau 测量 改进了 AD 和 MCI 与 CTL 的分类 在本文提出的研究中，我们重点关注 CSF 的效用。 EV 及其货物作为 AD 更强大的生物标志物，显示出女性与男性的明显差异， 具有 APOE-e4 与 APOE-e3 基因型的人 我们在人类脑脊液 EV 和死后大脑中的试点数据。 表明：i) AD 中的膜颗粒尺寸大于 CTL CSF；ii) CSF EV 具有表面标记； AD 和/或帕金森病 (PD) 特有的蛋白质与 CTL 相比；iii) CSF EV miRNA 有所改善； AD 与总 CSF 的预测能力；iv) CSF EV miRNA 显示性别和 APOE 依赖性变化 v) 与 CTL 相比，女性 AD CSF EV 中的 miR-16-5p 有所增加；以及 vi) 的预测基因靶标 miR-16-5p、SNAP-25 在女性 AD 海马中减少，这些研究共同支持了性别和 AD 之间的关系。 APOE-e4 都会导致 AD CSF EV 的变化，这可能是 AD 患病率较高的部分原因 女性 AD 以及 APOE-e4 基因型人群患 AD 的风险增加因此，我们建议。 在性别和性别背景下，建立 AD CSF EV 与 CTL 的物理特征和分子货物 我们将使用我们的信息论方法来识别最强的依赖性。 区分 AD、PD 和 CTL，并确定高风险 AD 的候选生物标志物。 女性 PD 识别 CSF EV 表面标记和 AD 特有的货物，然后我们将识别和识别。 验证与 AD 相关的 CSF EV miRNA 的基因靶点，我们的研究将共同​​确定具体的目标。 通过结合脑脊液中的物理特征、表面标记物谱、miRNA 和蛋白质来确定 AD 的生物标记物 他们还将在人脑中建立 AD EV miRNA 的神经相关基因靶标。 可能有助于 AD 发病机制的过程，并作为 AD 的新治疗靶点。