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.

项目摘要尽管对早期诊断阿尔茨海默氏病的兴趣浓厚，但面临许多基本细微差别和混合病理，进展受到不足的阻碍对病理的敏感性和低特异性。新的分子测定包括脑脊液通过正电子发射断层扫描（PET）测量和脑淀粉样菌斑成像。但是，这些技术要么需要腰穿，要么面临限制对早期检测的敏感性不足，或者对于更广泛的采用而言太昂贵了。因此，及时且具有成本效益的AD诊断有未满足的需求。可以原则上检测血液中疾病生物标志物，称为“液体活检” 提高测量几乎不可见疾病的准确性。虽然现有的液体活检涉及血液循环生物标志物的主要主要，在其他体液中检测到它们眼泪可能完全是非侵入性的。外泌体是细胞外的细胞外的含有表面蛋白质和遗传材料（DNA和RNA）的蔬菜，反映了父母细胞的特征和构成。分析脑源性外泌体（BDE）将因此，直接了解神经元和神经胶质起源的亲本细胞状态。为了尤其是最近的证据表明，阿尔茨海默氏病诊断尤其是病理表面蛋白和货物微RNA在AD中的BDE中的表达不同。因此，解锁循环bde中的大量信息可能会导致范式转移。但是，目前对bde进行分析的局限性如下：（1）只有血液是研究（2）复杂的协议；（3）无标签的尺寸/计数缺乏分子特异性；（4）提供高度平均的结果，具有正常外泌体的高背景，从而导致敏感性不佳。（5）提供“部分”信息：表面抗原或货物DNA/RNA，但不是两个。以上所有这些都阻碍了对BDE的进一步理解的发展与AD和AD诊断的潜在撕裂测试相关。在此行政补充中，我们建议延长微流体阵列的纳米质传感器和执行器（MANSA）原始R01中的平台以泪水为单一的BDE分析。 MANSA平台将启用：（1）简化了从捕获到分析的操作。（2）通过数字计数提高灵敏度通过动态成像。（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。高分辨率的外泌体阵列表面蛋白质和货物microRNA剖面将有助于高动态范围枚举和在基于泪水的AD诊断中提高灵敏度，并强调高价值AD生物标志物。这拟议的技术将导致一种经济高效的，对护理友好的，翻译的平台。