Streamlined and comprehensive brain-derived tear exosome profiling by microfluidic arrayed nanoplasmonic sensors and actuators

通过微流体阵列纳米等离子体传感器和执行器进行简化和全面的脑源性泪液外泌体分析

基本信息

批准号：
10712272
负责人：
Wei-Chuan Shih
金额：
$ 15.5万
依托单位：
UNIVERSITY OF HOUSTON
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：
2021-06-15 至 2025-03-31
项目状态：
未结题

项目摘要

PROJECT SUMMARY Despite keen interests in early diagnosis of Alzheimer’s disease, which faces a multitude of underlying nuanced and mixed pathologies, the progress has been hampered by insufficient sensitivity and low specificity to the pathology. New molecular assays include cerebrospinal fluid measurements and brain amyloid plaque imaging through positron emission tomography (PET). However, these techniques face limitations as they either require lumbar punctures, have insufficient sensitivity for early detection, or are too expensive for wider adoption. As a result, there is an unmet need in timely and cost-effective AD diagnostics. Detection of disease biomarkers in the blood, known as “liquid biopsy”, can in principle improve the accuracy of measuring nearly invisible diseases. Although existing liquid biopsy concerns primarily with blood circulating biomarkers, detecting them in other bodily fluids such as tears would potentially be completely non-invasive. Exosomes are cell-excreted extracellular vesicles that contain surface proteins and genetic materials (DNA and RNA) that reflect the characteristics and make-up of the parental cells. Analyzing brain-derived exosomes (BDE) would therefore provide direct insight into the state of the parental cells of neuronal and glial origins. For Alzheimer’s disease diagnostics, in particular, recent pieces of evidence have shown that several pathological surface proteins and cargo micro-RNAs are differentially expressed in BDE in AD. Therefore, unlocking the wealth of information in circulating BDE can potentially cause a paradigm shift. However, current limitations for profiling BDE are the following: (1) only blood has been studied; (2) sophisticated protocols; (3) label-free sizing/counting lacks molecular specificity; (4) providing highly averaged results with high background from normal exosomes, thus leading to poor sensitivity. (5) provide “partial” information: either surface antigen or cargo DNA/RNA, but not both. All of the above has hampered the development of further understanding of BDE associated with AD and a potential tear test for AD diagnostics. In this Administrative Supplement, we propose to extend the microfluidic arrayed nanoplasmonic sensor & actuator (MANSA) platform in the original R01 to single BDE profiling in tears. The MANSA platform will enable: (1) streamlined operation from capturing to profiling. (2) improved sensitivity by digital counting via dynamic imaging. (3) improved specificity by profiling multiple surface proteins and cargo microRNA associated with AD pathologies. A high-resolution, exosome array with multiplex surface protein and cargo microRNA profiles will facilitate high dynamic range enumeration and boost sensitivity in tear-based AD diagnostics and accentuate high-value AD biomarkers. The proposed technology would lead to a cost-effective, point-of-care-friendly, translational platform.

项目概要尽管人们对阿尔茨海默氏病的早期诊断抱有浓厚的兴趣，但该病面临着许多挑战由于潜在的微妙和混合的病症，进展受到了不足的阻碍新的分子检测包括脑脊液，其敏感性和特异性较低。通过正电子发射断层扫描（PET）进行测量和脑淀粉样斑块成像。然而，这些技术面临局限性，因为它们要么需要腰椎穿刺，要么需要早期检测的灵敏度不够，或者对于更广泛的采用来说过于昂贵。对及时且具有成本效益的 AD 诊断的需求尚未得到满足。血液中疾病生物标志物的检测（称为“液体活检”）原则上可以尽管现有液体活检，但提高了测量几乎看不见的疾病的准确性。主要关注血液循环生物标志物，在其他体液中检测它们，例如眼泪可能是完全非侵入性的。含有表面蛋白和遗传物质（DNA 和 RNA）的囊泡，反映了分析脑源性外泌体（BDE）将因此，可以直接了解神经和神经胶质起源的亲本细胞的状态。阿尔茨海默病的诊断，特别是最近的证据表明，一些 AD 中 BDE 中病理表面蛋白和货物 micro-RNA 的表达存在差异。因此，释放流通中的 BDE 中的大量信息可能会引发一种范式然而，目前 BDE 分析的局限性如下：(1) 仅对血液进行了分析。研究；（2）复杂的方案；（3）无标记的大小/计数缺乏分子特异性；提供具有正常外泌体高背景的高度平均结果，从而导致 (5) 提供“部分”信息：表面抗原或货物DNA/RNA，但是但并非两者都阻碍了对 BDE 的进一步了解。与 AD 相关以及用于 AD 诊断的潜在撕裂测试。我们建议扩展微流体阵列纳米等离子体传感器和执行器（MANSA）原始 R01 平台中的单一 BDE 剖析眼泪 MANSA 平台将启用： (1) (2) 通过数字计数提高灵敏度 (3) 通过分析多种表面蛋白和货物来提高特异性。与 AD 病理相关的 microRNA。表面蛋白和货物 microRNA 图谱将促进高动态范围计数和提高基于泪液的 AD 诊断的灵敏度，并强调高价值的 AD 生物标志物。拟议的技术将带来具有成本效益、护理点友好、可转化的技术平台。