Single Exosome Technology for Alzheimer's Disease

单一外泌体技术治疗阿尔茨海默病

• 批准号: 10330840
• 负责人: GREGORY W FARIS
• 金额: $ 29.92万
• 依托单位: NUMENTUS TECHNOLOGIES INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10330840
• 关键词: Address; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease diagnosis; Alzheimer' s disease diagnostic; Alzheimer' s disease pathology; Alzheimer’s disease biomarker; Amyloid beta-Protein; Amyotrophic Lateral Sclerosis; Area; Autopsy; Biological; Biological Assay; Biological Markers; Blood; Blood - brain barrier anatomy; Blood specimen; Brain; Cardiovascular Diseases; Cell Culture Techniques; Cells; Centrifugation; Cerebrospinal Fluid; Clinical; Clinical Management; Cognitive; Consensus; Cost Analysis; Data; Detection; Diagnosis; Diagnostic; Diagnostics Research; Disease; Disease Progression; Elderly; Evaluation; Funding; Gender; Health; Hematological Disease; Hour; Human; Image; Immunoprecipitation; Individual; Inflammation Mediators; Lead; Malignant Neoplasms; Measures; Mediator of activation protein; Membrane Proteins; Messenger RNA; Methods; MicroRNAs; NCAM1 gene; Neural Cell Adhesion Molecule L1; Neuraxis; Neurodegenerative Disorders; Neurologic; Neurons; Nucleic Acids; Outcome; Parkinson Disease; Pathology; Patients; Peripheral; Phase; Physiology; Population; Preparation; Procedures; Process; Prognosis; Proteins; Reporting; Research; Research Institute; Sampling; Sensitivity and Specificity; Signal Transduction; Source; Speed; Spinal Puncture; Structure of superior frontal gyrus; Surface; Technology; Testing; The Sun; Time; Untranslated RNA; Venous blood sampling; amplification detection; anandamide; base; design; detection method; exosome; expectation; extracellular vesicles; fluorescence imaging; imaging platform; imaging system; improved; innovation; liquid biopsy; mild cognitive impairment; minimally invasive; nanoparticle; nervous system disorder; neuroinflammation; new technology; novel; particle; peripheral blood; prognostic tool; screening; systems research; tau Proteins; tau-1; virtual

PROJECT SUMMARY This project responds to the important need for improved diagnostics for Alzheimer’s disease (AD). Existing methods based on biomarkers such as Ab and tau protein ratios from cerebrospinal fluid (CSF) are useful but incomplete. Furthermore, CSF sampling requires lumbar puncture and is too expensive for broad screening. A class of extracellular vesicles (EVs), exosomes, provide an attractive target for AD diagnostics. Exosomes freely cross the blood-brain barrier and can be readily sampled in peripheral blood, enabling a blood-based liquid biopsy. Exosomes also provide rich signatures for disease detection, including both proteins and nucleic acids (mRNA, miRNA, and other non-coding RNAs). The majority of studies on EVs and AD have been performed with bulk or batch analyses. Bulk analysis has a fundamental limitation because the relatively rare exosomes specific to the central nervous system (CNS) are easily confounded (swamped) by the exosome contributions of peripheral cells. To overcome this limitation, we are developing methods for combined protein/nucleic acid analysis in single exosomes. Our innovative, high-content, high-throughput method is designed to simultaneously analyze, in one pass, up to 10 potential AD biomarker cargoes in as many as 107 individual, CNS-tagged blood exosomes. These unique capabilities provide multiple advantages over previous approaches. Our method can rapidly: 1) distinguish and separately analyze both exosomes and other EVs; 2) discriminate and simultaneously evaluate multiple CNS-specific exosome surface markers, whereas conventional approaches can only evaluate one CNS-specific surface marker at a time, significantly limiting the ability to identify exosomes of CNS origin; 3) individually interrogate each and every exosome in a sample for its cargoes, dramatically raising information content compared to conventional methods where exosome cargoes must be pooled; and 4) search for unique combinations of biomarkers within unique, CNS-specific exosomal populations, an impossibility with conventional approaches. We therefore propose the following stepwise objectives for this Phase I project. First, to optimize the combined protein/nucleic acid analysis of exosomes produced by human SH-SY5Y cells. Second, to assay exosomes in brain homogenate samples from rapid autopsies of 40 AD, 40 mild cognitive impairment (MCI), 40 non-AD neurological conditions (nADneuro) (e.g., Parkinson’s disease, amyotrophic lateral sclerosis), and 40 normal elderly control (NC) subjects. Third, to assay exosomes in rapid autopsy blood samples from AD, MCI, nADneuro, and NC subjects that provided brain samples used in Objective 2. We anticipate that our novel imaging platform has the potential to become a new research/diagnostic/ prognostic tool for the clinical management of AD or other pathologies in which EV/exosomal analysis could provide clinically useful information, such as other neurodegenerative diseases, cancer, and cardiovascular disease. This capability may even assist in developing exosome-based therapies for these pathologies.

项目摘要 该项目应对改善阿尔茨海默氏病（AD）诊断的重要需求。现存的 基于脑脊液（CSF）的AB和TAU蛋白比等生物标志物的方法很有用，但 不完整。此外，CSF采样需要腰椎穿刺，对于广泛的筛查来说太昂贵了。一个 细胞外蔬菜（EVS）类别，外泌体为AD诊断提供了有吸引力的靶标。外泌体 自由地越过血脑屏障，可以轻松地在外围血液中取样，使血液基于血液 液体活检。外泌体还为疾病检测提供了丰富的特征，包括蛋白质和核酸 酸（mRNA，miRNA和其他非编码RNA）。关于电动汽车和AD的大多数研究是 通过批量或批处理分析进行。批量分析具有基本限制，因为相对罕见 特定于中枢神经系统（CNS）的外泌体很容易被外泌体混淆（淹没） 外围细胞的贡献。为了克服这一限制，我们正在开发合并的方法 单个外泌体中的蛋白质/核酸分析。我们的创新，高素质，高通量方法是 旨在同时在一次通道中分析多达107个潜在的AD生物标志物货物多达107个货物 个体，CNS标记的血液外泌体。这些独特的功能比以前具有多个优势 方法。我们的方法可以迅速：1）独特和分别分析外泌体和其他电动汽车； 2） 区分并简单地评估多个CNS特异性外泌体表面标记，而 常规方法只能一次评估一个CNS特异性的表面标记，从而显着限制 能够鉴定中枢神经系统起源的外泌体； 3）单独询问样本中的每个外泌体 它的货物，与传统方法相比，大幅度提高了信息含量 必须合并货物； 4）在独特的，特定于中枢神经系统的特定于独特的生物标志物中搜索独特的组合 外泌体种群，对常规方法的不可能。因此，我们提出以下内容 该阶段项目的逐步目标。首先，优化蛋白质/核酸的组合分析 人类SY5Y细胞产生的外泌体。第二，在脑匀浆样品中断言外泌体 从40 AD的快速尸检，40个轻度认知障碍（MCI），40个非AD神经系统条件 （Nadneuro）（例如，帕金森氏病，肌萎缩性侧索硬化症）和40个正常对照（NC） 主题。第三，在AD，MCI，Nadneuro和NC受试者的快速尸检血液样本中断言外泌体 这提供了目标2中使用的大脑样品。 我们预计我们的新颖成像平台有可能成为一项新的研究/诊断/ 用于EV/外泌体分析的AD或其他病理的临床管理的预后工具可以 提供临床上有用的信息，例如其他神经退行性疾病，癌症和心血管 疾病。这种能力甚至可以帮助为这些病理发展开发基于外泌体的疗法。