Brain-derived extracellular vesicles as a novel source of biomarkers for disease progression and environmental exposure in ALS

脑源性细胞外囊泡作为 ALS 疾病进展和环境暴露生物标志物的新来源

• 批准号: 9887409
• 负责人: Diane Berengere Re
• 金额: $ 53.93万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9887409
• 关键词: ALS patients; Amyotrophic Lateral Sclerosis; Astrocytes; Autopsy; Back; Biological Markers; Blood; Blood Circulation; Brain; Cells; Cerebrospinal Fluid; Clinical; Collection; Communication; Disease; Disease Progression; Dose; Elements; Environmental Exposure; Enzyme-Linked Immunosorbent Assay; Etiology; Evaluation; Exposure to; GLAST Protein; Hair; Human; In Vitro; Lead levels; Link; Longitudinal Studies; Measurement; Measures; Mediating; Membrane; Metal exposure; Metals; MicroRNAs; Molecular Profiling; Motor Neurons; Neural Cell Adhesion Molecule L1; Neuraxis; Neuroglia; Neurons; Nucleic Acids; Organ; Paralysed; Pathogenicity; Pathway interactions; Patients; Peripheral; Phenotype; Play; Prevention; Proteins; Reporting; Research; Role; Sampling; Signal Transduction; Son of Sevenless Proteins; Source; Surface; System; Testing; Time; Tissues; Toxic Environmental Substances; Travel; Treatment Efficacy; Tube; Veterans; Visit; base; biobank; capsule; cohort; effective therapy; extracellular vesicles; improved; miRNA expression profiling; minimally invasive; motor neuron degeneration; nervous system disorder; neurotoxic; novel; novel marker; patient biomarkers; protein TDP-43; public health relevance; toxic metal; transcriptome sequencing

SUMMARY Amyotrophic lateral sclerosis (ALS) is a mostly sporadic and invariably fatal paralytic disorder. ALS pathogenic mechanisms are elusive and its etiology is poorly understood, even if environmental exposures, including toxic metals, are thought to play a key role. Progresses in both therapy and prevention for ALS have been held back by the lack of clear biomarkers, indicative of patients' disease progression and central nervous system (CNS) exposure to environmental toxicants. Here, we propose to test whether the cargoes of blood extra-cellular vesicles (EVs), i.e., tiny membrane-bound capsules that shuttle biomolecules out of the CNS, can serve as novel biomarkers of CNS metal exposure and disease progression in ALS. EVs carry nucleic acids (including microRNAs [miRNAs]), proteins, and other elements, while circulating throughout the body for inter- organ communication. EVs also operate as "trash bags" allowing cells to eliminate excesses of unwanted cellular materials, including toxic metals and proteins. CNS-EVs can be isolated from blood because they express on their surface the neuron-specific protein L1CAM or the astrocyte-specific protein GLAST. Blood CNS-EVs could open a direct window of observation into the ALS brain from the periphery and provide different types of biomarkers. First, CNS-EVs may reflect disease progression by carrying pathogenic proteins progressively accumulating in neurons and astrocytes in ALS, such as the TAR DNA-binding protein 43 (TDP-43), or by showing progressive changes in miRNA profiles. Second, CNS-EV metal levels could be a direct surrogate of CNS metal load, and as suggested by metals' effects on miRNA profiles, they could provide a molecular fingerprint of patient metal exposure, when direct measurement is not possible. To test these hypotheses, we will leverage unique biospecimen collections from three well-phenotyped cohorts of ALS patients and a set of 140 control samples: (1) the COSMOS multi-center cohort (n=269 patients; 2-6 serial visits; blood); (2) the NEALS longitudinal study for ALS biomarkers (LABB) (N=150) with 2-3 visits; and (3) the Veterans Affairs Biorepository (VAB) (n=147; paired blood and postmortem CNS tissues). Clinical stage was assessed at each blood draw through the ALS Functional Rating Scale-Revised. Our specific aims are to: Aim-1: Test whether TDP-43 level in blood L1CAM/GLAST-EVs is a biomarker of ALS progression by ELISA quantification of TDP-43 in the serial COSMOS and LABB blood draws, and testing its association with ALS progression. Aim- 2: Assess metal content in blood CNS-EV as a biomarker of patient's exposure by measuring the levels of 7 metals linked to ALS in paired CNS-EVs and CNS tissues (VAB), and testing their association with ALS progression (LABB). Aim-3: Test whether miRNA profiles in blood CNS-EVs are biomarkers of both ALS progression and metal exposure by miRNA-sequencing and assessing whether they mediate TDP-43's and metals' effect on ALS progression. The novel and minimally invasive EV biomarkers tested here could greatly improve the evaluation of ALS treatment efficacy, and revolutionize metal exposure assessment.

概括 肌萎缩侧索硬化症 (ALS) 是一种散发性且总是致命的麻痹性疾病。 ALS致病性 即使环境暴露，包括有毒物质，其机制仍难以捉摸，其病因也知之甚少。 金属被认为发挥着关键作用。 ALS 的治疗和预防进展均受到阻碍 由于缺乏明确的生物标志物来指示患者的疾病进展和中枢神经系统（CNS） 接触环境毒物。在这里，我们建议测试血液中的细胞外货物是否 囊泡（EV），即将生物分子运送出中枢神经系统的微小膜结合胶囊，可以充当 中枢神经系统金属暴露和 ALS 疾病进展的新型生物标志物。 EV携带核酸（包括 microRNA [miRNA]）、蛋白质和其他元素，同时在全身循环以进行器官间 沟通。电动汽车还充当“垃圾袋”，允许细胞消除多余的不需要的细胞 材料，包括有毒金属和蛋白质。 CNS-EV 可以从血液中分离出来，因为它们表达于 它们的表面有神经元特异性蛋白 L1CAM 或星形胶质细胞特异性蛋白 GLAST。血液 CNS-EV 可以打开从外围直接观察 ALS 大脑的窗口，并提供不同类型的 生物标志物。首先，CNS-EVs可能通过逐渐携带致病蛋白来反映疾病进展 在 ALS 中的神经元和星形胶质细胞中积累，例如 TAR DNA 结合蛋白 43 (TDP-43)，或通过 显示 miRNA 图谱的渐进变化。其次，CNS-EV 金属水平可以直接替代 CNS 金属负载，正如金属对 miRNA 图谱的影响所表明的，它们可以提供分子 当无法直接测量时，患者金属暴露的指纹。为了检验这些假设，我们 将利用来自三个表型良好的 ALS 患者队列和一组 140 个对照样本：(1) COSMOS 多中心队列（n=269 名患者；2-6 次连续访视；血液）； （2） NEALS 针对 ALS 生物标志物 (LABB) 的纵向研究 (N=150)，进行 2-3 次访视； (3) 退伍军人事务部 生物储存库 (VAB)（n=147；配对血液和死后中枢神经系统组织）。每个阶段均评估临床阶段 通过 ALS 功能评定量表（修订版）抽血。我们的具体目标是： Aim-1：测试是否 血液 L1CAM/GLAST-EV 中的 TDP-43 水平是通过 ELISA 定量的 ALS 进展的生物标志物 连续 COSMOS 和 LABB 抽血中的 TDP-43，并测试其与 ALS 进展的关联。目的- 2：通过测量 CNS-EV 的水平来评估血液 CNS-EV 中的金属含量，作为患者暴露的生物标志物 配对 CNS-EV 和 CNS 组织 (VAB) 中与 ALS 相关的 7 种金属，并测试它们与 ALS 的关联 进展（LABB）。 Aim-3：测试血液 CNS-EV 中的 miRNA 谱是否是两种 ALS 的生物标志物 通过 miRNA 测序确定进展和金属暴露，并评估它们是否介导 TDP-43 和 金属对 ALS 进展的影响。这里测试的新型微创 EV 生物标志物可以极大地 提高 ALS 治疗效果的评估，彻底改变金属暴露评估。