Automated High-purity Exosome isolation-based AD diagnostics system (AHEADx)

基于自动化高纯度外泌体分离的 AD 诊断系统 (AHEADx)

• 批准号: 10738697
• 负责人: Tony Jun Huang
• 金额: $ 78.88万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738697
• 关键词: Acceleration; Acoustics; Address; Affect; Alzheimer disease detection; Alzheimer disease screening; Alzheimer' s Disease; Alzheimer' s disease diagnosis; Alzheimer' s disease diagnostic; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease test; Alzheimer' s disease therapeutic; Alzheimer' s disease therapy; Alzheimer’s disease biomarker; Behavioral; Benchmarking; Biological Markers; Biometry; Blood; Blood - brain barrier anatomy; Brain Pathology; Brain imaging; Cerebrospinal Fluid; Clinical; Clinical Research; Communication; Computer software; Detection; Development; Devices; Diagnosis; Diagnostic Procedure; Diagnostic tests; Disease; Disease Progression; Early Diagnosis; Early treatment; Economic Burden; Electronics; Future; Healthcare; Impairment; Intuition; Language; Libraries; Liquid substance; Longitudinal Studies; Machine Learning; Memory impairment; Methods; MicroRNAs; Molecular; Monitor; NanoPillar; Neurodegenerative Disorders; Nucleic Acids; Optics; Patient-Focused Outcomes; Patients; Performance; Persons; Physiological; Plasma; Positron-Emission Tomography; Proteins; Research; Research Personnel; Risk; Sampling; Source; Specificity; Speech; Speed; Spinal Puncture; Standardization; Symptoms; System; Techniques; Technology; Testing; Therapeutic; Time; United States; Universities; Validation; Whole Blood; accurate diagnosis; amplification detection; biomaterial compatibility; clinical application; clinical diagnosis; cognitive ability; cost; cost efficient; diagnostic platform; diagnostic tool; exosome; extracellular vesicles; functional decline; global health; health economics; imaging modality; improved; invention; liquid biopsy; medical schools; minimally invasive; nanophotonic; nanoplasmonic; nanoscale; novel; photonics; plasmonics; point of care; point-of-care diagnostics; pre-clinical; prototype; response; screening; tool; trafficking; treatment response; treatment strategy

PROJECT SUMMARY Alzheimer’s disease (AD) is a severe neurodegenerative illness that destroys cognitive abilities causing memory impairment, difficulties with speech and language, behavioral changes, functional decline, and significant impairment. AD affects an estimated 13.8 million people in the United States and 50 million worldwide, imposing a significant economic burden and global health crisis. While many researchers are working towards developing a cure for the disease, there is currently no objective, point-of-care diagnostic test for the early diagnosis of AD, significantly limiting screening efforts for clinical studies and delaying treatment for patients suffering from early AD pathology. The current standard methods for AD diagnosis involve expensive PET imaging methods that irradiate the patient and cerebrospinal fluid biomarker tests, which are invasive and require a lumbar puncture. Recently, exosomes (30-150 nm extracellular vesicles) have been identified as a possible tool for AD diagnosis, attributed to their ubiquitous presence in biofluids, their ability to pass through the blood-brain-barrier, and their rich library of AD-relevant physiological information present in the molecular cargo they carry, making them prime candidates for use as biomarkers. However, the clinical application of exosomes is hindered by slow, inefficient techniques for exosome isolation and the absence of standardized exosomal biomarker detection and analysis. Thus, an automated, highly sensitive, fast, and efficient system that can isolate exosomes from biofluids and analyze the miRNAs and proteins they contain will significantly improve early-stage AD diagnosis efforts. In this R01 project, we will develop an Automated High-purity Exosome isolation-based AD diagnostics system (AHEADx) to address the limitations of current technologies. The proposed AHEADx platform includes two units: (1) a rapid (<1 min), high-yield (>90%), and high-purity (>90%) acoustic Bessel beam-based separation unit to isolate and enrich exosomes from whole blood, and (2) a rapid (<6 mins), highly sensitive photonic PCR and immuno-PCR (~1 copy/µL for nucleic acids and ~5 copies/µL for proteins, respectively) utilizing a plasmonic nanopillar array to enable on-chip thermocycling and multiplexed exosomal screening of combined panels of AD-relevant biomarkers. Our rapid and precise AHEADx platform will provide a simple, minimally invasive liquid biopsy to detect molecular AD biomarkers with ultrahigh accuracy and sensitivity in early-stage AD patients, allowing for an effective diagnostic method of AD screening for earlier treatment before the onset of severe symptoms of the disease and enable long-term studies on AD development and progression. Additionally, the proposed technology will accelerate the discovery of new exosomal miRNA and protein AD biomarkers and help to elucidate the mechanisms in which exosome trafficking and transport contribute to AD pathology. With these advantages, our AHEADx platform can potentially exceed current clinical standards in AD diagnostics, addressing a significant need in the field and providing a compelling platform for earlier, accurate, and sensitive detection of AD and long-term studies on the effects of novel AD therapeutics.

项目总结 阿尔茨海默病(AD)是一种严重的神经退行性疾病，会破坏导致记忆的认知能力 功能障碍、言语和语言障碍、行为改变、功能衰退和显著 减损。据估计，美国有1380万人受到广告的影响，全球有5000万人受到广告的影响。 严重的经济负担和全球健康危机。当许多研究人员正在努力开发 作为一种治愈疾病的方法，目前还没有客观的、临床点诊断测试来早期诊断阿尔茨海默病， 大大限制了临床研究的筛查工作，并推迟了对早期糖尿病患者的治疗 公元病理学。目前诊断AD的标准方法包括昂贵的PET成像方法， 照射患者和脑脊液生物标记物测试，这是侵入性的，需要腰椎穿刺术。 最近，Exosome(30-150 nm细胞外小泡)已被确定为一种可能的AD诊断工具。 归因于它们在生物体液中的普遍存在，它们通过血脑屏障的能力，以及它们的 丰富的AD相关生理信息库存在于它们携带的分子货物中，使它们成为主要的 作为生物标志物的候选者。然而，外切体的临床应用受到了缓慢、低效的阻碍。 外切体分离技术和缺乏标准化的外切体生物标志物检测和分析。 因此，一种自动化、高度敏感、快速和高效的系统可以从生物体液中分离外切体和 分析它们所包含的miRNAs和蛋白质将显著改善早期AD诊断工作。在这 在R01项目的基础上，我们将开发一个基于自动化高纯度Exosome分离的AD诊断系统 (AHEADx)以解决当前技术的限制。拟议的AHEADx平台包括两个单元： (1)快速(&lt；1分钟)、高产量(&gt；90%)和高纯度(&gt；90%)的基于声学贝塞尔波束的分离单元 从全血中分离和浓缩外切体，以及(2)快速(&lt；6分钟)、高灵敏的光子聚合酶链式反应和 血浆免疫聚合酶链式反应(核酸~1拷贝/微米L，蛋白质~5拷贝/微米L) 纳米管阵列可实现芯片上热循环和多路外体筛选的组合面板 与广告相关的生物标志物。我们快速而精确的AHEADx平台将提供简单、微创的液体 以超高准确度和灵敏度检测早期AD患者的分子AD生物标志物的活检 为重症发作前的早期治疗提供了一种有效的AD筛查诊断方法 这将有助于对阿尔茨海默病的症状进行研究，并对阿尔茨海默病的发展和进展进行长期研究。此外， 拟议的技术将加速发现新的外体miRNA和蛋白质AD生物标记物，并有助于 阐明外切体运输和运输在AD病理中的作用机制。有了这些 优势，我们的AHEADx平台在AD诊断方面可能会超过当前的临床标准， 满足现场的重大需求，并为更早、更准确、更敏感地提供一个引人注目的平台 阿尔茨海默病的检测和新的阿尔茨海默病治疗方法的长期研究。