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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10712272
• 关键词: Address; Administrative Supplement; Adoption; Alzheimer' s Disease; Alzheimer' s disease diagnosis; Alzheimer' s disease diagnostic; Alzheimer' s disease pathology; Alzheimer' s disease test; Alzheimer’s disease biomarker; Amyloid beta-Protein; Antibodies; Area; Binding; Biological Assay; Biological Markers; Blood; Body Fluids; Brain; Brain Pathology; Bypass; Cells; Cerebrospinal Fluid; Characteristics; Collaborations; DNA; Data; Data Analyses; Deposition; Detection; Development; Diagnostic; Diameter; Disease; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Event; Excretory function; Face; Fluorescent Dyes; Friends; Genetic Markers; Genetic Materials; Genomics; Goals; Health Sciences; Image; Label; Letters; Liquid substance; Measurement; Measures; Membrane Proteins; Methodology; MicroRNAs; Microbubbles; Microfluidics; Microscopic; Mission; Molecular; Molecular Profiling; Monitor; Neurons; Onset of illness; Pathologic; Pathology; Plasma; Play; Positron-Emission Tomography; Procedures; Proteins; Protocols documentation; Quantum Dots; RNA; Research; Resolution; Sampling; Senile Plaques; Signal Transduction; Specificity; Spinal Puncture; Stains; Surface; Surface Antigens; Surveys; Techniques; Technology; Testing; Texas; United States National Institutes of Health; Universities; Work; biochip; burden of illness; circulating biomarkers; cost; cost effective; density; design; detection limit; diagnostic assay; differential expression; digital; early onset; exosome; extracellular; extracellular vesicles; healthy volunteer; improved; indexing; insight; interest; liquid biopsy; miRNA expression profiling; microRNA biomarkers; multiplex assay; nanodisk; nanoplasmonic; new technology; novel strategies; operation; point of care; research and development; sensor; success; tau Proteins; technology platform; transcriptomics; volunteer

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 尽管对阿尔茨海默病的早期诊断有着浓厚的兴趣，但阿尔茨海默病面临着许多问题。 潜在的细微差别和混合的病理，进展一直受到阻碍， 对病理学的敏感性和低特异性。新的分子检测包括脑脊液 测量和通过正电子发射断层扫描（PET）的脑淀粉样斑块成像。 然而，这些技术面临局限性，因为它们要么需要腰椎穿刺， 对于早期检测来说灵敏度不足，或者对于更广泛的采用来说太昂贵。因此，在本发明中， 在及时和成本有效的AD诊断方面存在未满足的需求。 检测血液中的疾病生物标志物，被称为“液体活检”，原则上可以 提高测量几乎不可见疾病的准确性。虽然现有的液体活检 主要关注血液循环生物标志物，在其他体液中检测它们， 撕裂可能是完全无创的。外泌体是细胞分泌到细胞外的 含有表面蛋白质和遗传物质（DNA和RNA）的囊泡， 亲本细胞的特征和组成。分析脑源性外泌体（BDE）将 因此提供了对神经元和神经胶质起源的亲代细胞状态的直接洞察。为 特别是阿尔茨海默病的诊断，最近的证据表明， 病理性表面蛋白和货物微RNA在AD的BDE中差异表达。 因此，释放流通中的溴化二苯醚的丰富信息可能会导致一种范式， 班然而，目前对溴化二苯醚进行定性分析的局限性如下：（1）仅对血液进行了定性分析， 研究;（2）复杂的方案;（3）无标记大小测定/计数缺乏分子特异性;（4） 提供具有来自正常外来体的高背景的高度平均结果，从而导致 敏感性差。(5)提供“部分”信息：表面抗原或货物DNA/RNA，但 不能两者都有。这些都阻碍了对溴化二苯醚的进一步认识 与AD相关，以及用于AD诊断的潜在撕裂试验。在本行政补充中， 我们建议扩展微流控阵列纳米等离子体激元传感器和致动器（曼萨） 原始R 01中的平台与单一溴化二苯醚特征分析中的裂缝。曼萨平台将使：（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病理学相关的microRNA一种高分辨率的外泌体阵列， 表面蛋白和货物microRNA谱将促进高动态范围计数， 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。的 拟议的技术将导致一个具有成本效益的，护理点友好的，翻译 平台

项目成果

Exploring the synergy of radiative coupling and substrate undercut in arrayed gold nanodisks for economical, ultra-sensitive label-free biosensing.

• DOI: 10.1109/jsen.2021.3111125
• 发表时间: 2021
• 期刊: IEEE sensors journal
• 影响因子: 4.3
• 作者: Misbah I;Ohannesian N;Qiao Y;Lin SH;Shih WC
• 通讯作者: Shih WC