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.

项目摘要 阿尔茨海默氏病（AD）是一种严重的神经退行性疾病，破坏了导致记忆的认知能力 障碍，语言和语言难度，行为变化，功能下降和重大 损害。广告影响美国估计有1,380万人，在全球范围内有5000万人 巨大的经济负担和全球健康危机。许多研究人员正在努力发展 治愈该疾病，目前尚无对AD早期诊断的客观，护理诊断测试， 显着限制了临床研究的筛查工作，并延迟了早期患者的治疗 广告病理学。当前的AD诊断标准方法涉及昂贵的PET成像方法 照射患者和脑脊液生物标志物测试，这些测试具有侵入性，需要腰椎穿刺。 最近，外泌体（30-150 nm细胞外蔬菜）被确定为AD诊断的可能工具， 归因于它们在生物流体中无处不在的存在，他们的通过血脑屏障的能力及其 丰富的广告相关物理信息库，它们携带的分子货物中存在 候选人用作生物标志物。但是，外泌体的临床应用受到缓慢，效率低下的阻碍 外泌体分离的技术以及缺乏标准化外泌体生物标志物检测和分析的技术。 那是一种自动化，高度敏感，快速和高效的系统，可以将外泌体与生物流体和 分析它们所含的miRNA和蛋白质将显着改善早期AD诊断工作。在这个 R01项目，我们将开发一个自动化的高纯度外部隔离隔离诊断系统 （提前X）解决当前技术的局限性。拟议的TearX平台包括两个单元： （1）快速（<1分钟），高收益（> 90％）和高纯度（> 90％）的声学贝塞尔束基分离单元 分离并富集全血的外泌体，（2）快速（<6分钟），高度敏感的光子PCR和 免疫PCR（核酸的〜1拷贝/µL，分别用于蛋白质的〜5拷贝/µl）使用血浆骨蛋白） 纳米阵列可实现片上热环和多重外泌体筛查的组合板的组合面板 与广告相关的生物标志物。我们的快速而精确的前面X平台将提供简单的，微创的液体 在早期AD患者中检测具有超高准确性和灵敏度的分子AD生物标志物的活检， 允许在严重发作之前采用有效的AD筛查诊断方法进行早期治疗 该疾病的症状并能够长期研究AD的发展和进展。另外， 拟议的技术将加速发现新的外泌体miRNA和蛋白质AD生物标志物，并帮助 阐明外泌体运输和运输有助于AD病理学的机制。与这些 优势，我们的前进平台可能会超过广告诊断中当前的临床标准， 满足该领域的巨大需求，并为早期，准确和敏感的平台提供引人注目的平台 检测AD和关于新型AD疗法影响的长期研究。