Project Goals: This project involves developing a point-of-care platform to detect circulating endothelial cells for diagnosing or monitoring vascular disease or injury.
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.
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.
