A Microfluidic Platform for Detecting Circulating Endothelial Cells at the Point-of-Care

用于在护理点检测循环内皮细胞的微流体平台

• 批准号: 1509921
• 负责人: Lydia Sohn
• 金额: $ 36万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1509921&HistoricalAwards=false
• 关键词: Microfluidic; Platform; Detecting; Circulating; Endothelial

Proposal Title: A Microfluidic Platform for Detecting Circulating Endothelial Cells at the Point-of-CareProject Goals: This project involves developing a point-of-care platform to detect circulating endothelial cells for diagnosing or monitoring vascular disease or injury.Nontechnical Abstract: Circulating endothelial cells (CECs) are indicators of vascular injury and/or disease. Clinical studies have shown that individuals with vascular disorders, such as ischemia, vascular trauma, acute myocardial infarction, sickle-cell anemia, vasculitis, pulmonary hypertension, and deep-vein thrombosis, have higher levels of CECs than healthy controls (who had no little to no CECs). Importantly, the number of CECs strongly correlated with the severity of injury or disease. Thus, CECs have diagnostic and prognostic importance. Current methods to detect CECs are overall inadequate, as they lack sensitivity, require a highly trained physician-scientist to interpret results, and cannot be performed in a physician's office. This project will develop a microfluidic platform that can detect CECs in patient blood at the point-of-care, enabling a physician to diagnose and monitor a patient's vascular disease or injury and also to respond quickly in acute cases. Beyond the obvious high medical/clinical impact, this project will have strong societal impact, as workshops will be held to demonstrate to students how different disciplines in engineering and science can come together to address real and important problems, just as had done for this medical project, and how students are capable of both engineering and scientific thinking.Technical Abstract: The overarching goal of this project is to develop a novel label-free microfluidics platform that screens for circulating endothelial cells (CECs). CECs are indicators of vascular injury and/or disease and are either shed from the vascular wall (mature CECs) or recruited from the bone marrow (endothelial progenitor cells or EPCs). Representing 0.01% to 0.0001% of the total mononuclear cells in peripheral blood, CECs are challenging to detect. The platform to be developed will utilize inertial fluid dynamics within a contraction-expansion array (CEA) to isolate candidate CECs from whole blood based on size. The platform will then screen the isolated cells using Node-Pore Sensing (NPS) to not only identify CECs from white blood cells but also to differentiate mature CECs from EPCs based on specific phenotypic profiles. NPS measures the transit time of a cell as it interacts (specifically or non-specifically) with antibodies functionalized in a microfluidic channel that has been segmented by nodes. Specific interactions between cell-surface receptors and the functionalized antibody retard the cell, leading to longer transit times and subsequent determination of a particular surface-marker presence. Overall, having the ability to identify and distinguish between mature CECs and EPCS in patient blood at the point-of-care would enable a physician to diagnose and monitor a patient's vascular disease or injury and also respond quickly in cases such as an acute myocardial infarction.This three-year project has three specific aims:-Aim 1: To optimize a contraction-expansion array (CEA) device such that it enriches CECs from whole blood. The CEA device will rely upon inertial forces to fractionate blood and CECs based on size. The device will be optimized for 100% recovery of cancer cells.-Aim 2: To expand the capabilities of NPS to screen for mature CECs and EPCS. A unique NPS platform with optimized coding and processing (optimum Barker codes, matched filtering, sparse deconvolution, linear classifiers, all of which are inspired by radar and telecommunications theory) will be designed and developed to enable high-resolution detection and classification of CECs.-Aim 3: To integrate the optimized NPS with the CEA device and also include a sorter downstream. The CEA device with NPS will be integrated onto a single platform, enabling isolation, analysis, and sorting of CECs from peripheral blood.

提案标题：一个微流体平台，用于检测皮关点目标的循环内皮细胞：该项目涉及开发一个护理平台，以检测用于诊断或监测血管疾病或损伤的循环内皮细胞。无内科技术摘要：循环的内皮细胞（CECS）（cecs）是疾病和/或/或/或/或/或/或/或/或/或/或/或/或/或或或或或或或或或一位或/或/或/或/或/或/或/或/或或/或或或或或或或或一位或/或/或/或/或/或/或/或或/或或或或或或或或或一位为/所/所示所致。 临床研究表明，血管疾病的患者，例如缺血，血管创伤，急性心肌梗塞，镰状细胞贫血，血管炎，肺动脉高压和深vein血栓形成，比健康对照（他们没有CECS）更高的CEC水平。重要的是，CEC的数量与伤害或疾病的严重程度密切相关。因此，CEC具有诊断和预后的重要性。 当前检测CEC的方法总体上是不足的，因为它们缺乏灵敏度，需要训练有素的医师科学家来解释结果，并且不能在医师的办公室中进行。 该项目将开发一个微流体平台，该平台可以在医疗点检测患者血液中的CEC，从而使医生能够诊断和监测患者的血管疾病或受伤，并在急性病例中迅速反应。 Beyond the obvious high medical/clinical impact, this project will have strong societal impact, as workshops will be held to demonstrate to students how different disciplines in engineering and science can come together to address real and important problems, just as had done for this medical project, and how students are capable of both engineering and scientific thinking.Technical Abstract: The overarching goal of this project is to develop a novel label-free microfluidics platform that screens for circulating endothelial cells （CEC）。 CEC是血管损伤和/或疾病的指标，要么是从血管壁（成熟CEC）脱落的，要么是从骨髓（内皮祖细胞或EPC）中募集的。 CEC代表周围血液中单核细胞总细胞的0.01％至0.0001％，检测到具有挑战性。要开发的平台将利用收缩膨胀阵列（CEA）中的惯性流体动力学，以根据大小将候选CEC与全血分离。然后，该平台将使用节点孔传感（NP）筛选孤立的细胞，不仅鉴定了来自白细胞的CEC，而且还基于特定的表型特征将成熟的CEC与EPC区分开。 NPS测量细胞的运输时间（特异性或非特异性）与在微流体通道中官能化的抗体相互作用，该抗体已被节点分割。细胞表面受体与功能化抗体之间的特定相互作用延伸细胞，从而导致更长的过境时间，并随后确定特定的表面标志物存在。 Overall, having the ability to identify and distinguish between mature CECs and EPCS in patient blood at the point-of-care would enable a physician to diagnose and monitor a patient's vascular disease or injury and also respond quickly in cases such as an acute myocardial infarction.This three-year project has three specific aims:-Aim 1: To optimize a contraction-expansion array (CEA) device such that it enriches CECs from whole blood. CEA设备将根据大小依靠惯性力来分割血液和CEC。该设备将被优化，以100％恢复癌细胞。-AIM 2：扩大NP的功能以筛选成熟的CEC和EPC。具有优化编码和处理的独特NPS平台（最佳的吠叫代码，匹配的过滤，稀疏的反卷积，线性分类器，所有这些都受到雷达和电信理论的启发），以启用高分辨率检测和分类CEC.-IAM 3：将最佳的NPS与CEA的NPS相结合，并将其包括在内，并将其集成到Sorters设备中，并将其包括在内。带有NP的CEA设备将集成到单个平台上，从而可以从外周血对CEC进行隔离，分析和分类。