Single Exosome Technology for Alzheimer's Disease

单一外泌体技术治疗阿尔茨海默病

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease diagnosis Alzheimer&apos s disease diagnostic Alzheimer&apos 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

项目摘要

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.

项目概要该项目满足了改进现有阿尔茨海默病 (AD) 诊断的重要需求。基于脑脊液 (CSF) 中 Ab 和 tau 蛋白比率等生物标志物的方法是有用的，但此外，脑脊液取样需要腰椎穿刺，对于广泛筛查来说费用太高。一类细胞外囊泡 (EV) 外泌体为 AD 诊断提供了一个有吸引力的目标。可以自由穿过血脑屏障，并且可以很容易地在外周血中取样，从而实现基于血液的检测液体活检还为疾病检测提供丰富的特征，包括蛋白质和核酸。大多数关于 EV 和 AD 的研究都是基于 mRNA、miRNA 和其他非编码 RNA。进行批量或批量分析有一个根本性的限制，因为这种分析相对较少。中枢神经系统（CNS）特异性的外泌体很容易被外泌体混淆（淹没）为了克服这一限制，我们正在开发组合方法。我们的创新、高含量、高通量方法是单个外泌体中的蛋白质/核酸分析。旨在一次性同时分析多达 107 种潜在 AD 生物标志物货物单独的 CNS 标记的血液外泌体比以前具有多种优势。我们的方法可以快速：1）区分并单独分析外泌体和其他 EV；2）区分并同时评估多个 CNS 特异性外泌体表面标记，而传统方法一次只能评估一种中枢神经系统特异性表面标记物，这极大地限制了识别 CNS 来源的外泌体的能力；3) 单独询问样本中的每个外泌体与传统方法相比，外泌体显着提高了信息内容货物必须集中；4) 在独特的、中枢神经系统特异性的生物标志物中寻找独特的组合因此，我们提出以下建议：该第一阶段项目的逐步目标首先是优化蛋白质/核酸的组合分析。其次，脑匀浆检测样品中的外泌体。来自对 40 例 AD、40 例轻度认知障碍 (MCI)、40 例非 AD 神经系统疾病的快速尸检 (nADneuro)（例如帕金森病、肌萎缩侧索硬化症）和 40 名正常老年人对照 (NC) 第三，检测来自 AD、MCI、nADneuro 和 NC 受试者的快速尸检血液样本中的外泌体。提供了目标 2 中使用的大脑样本。我们预计我们的新型成像平台有潜力成为新的研究/诊断/ 用于 AD 或其他病理学临床管理的预后工具，其中 EV/外泌体分析可以提供临床有用的信息，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能有助于开发针对这些疾病的外泌体疗法。