Node-Pore Sensing for Cellular Screening

用于细胞筛查的节点孔传感

• 批准号: 8893816
• 负责人: Michael Lustig
• 金额: $ 22.44万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8893816
• 关键词: Antibodies; Biological Sciences; Biomedical Research; Bone Marrow; Breast Cancer cell line; CD44 gene; Cell Separation; Cell Surface Receptors; Cell surface; Cells; Classification; Clinical; Code; Color; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease remission; Endothelial Cells; Epidermal Growth Factor Receptor; Flow Cytometry; Fluorochrome; Gene Expression; Goals; Health; Hematologic Neoplasms; Label; MCF10A cells; MCF7 cell; MDA MB 231; Malignant Neoplasms; Measures; Metastatic breast cancer; Methods; Microfluidics; Molecular; Monitor; Mucin-1 Staining Method; Neoplasm Circulating Cells; Noise; Patients; Population; Process; Radar; Research; Residual Neoplasm; Resolution; Scientist; Signal Transduction; Solid Neoplasm; Sorting - Cell Movement; Surface; System; TACSTD2 gene; Techniques; Technology; Telecommunications; Time; base; cancer cell; cell population study; clinical Diagnosis; clinical application; cost; design; high reward; high risk; high throughput screening; improved; innovation; mechanical pressure; novel; peripheral blood; point of care; point-of-care diagnostics; regenerative; research study; screening; stem cell differentiation; stem cell population; theories

DESCRIPTION: The overarching goal of this project is to develop Node-Pore Sensing (NPS), an innovative label-free microfluidic technique, such that it could go beyond flow cytometry in terms of number of markers that can be practically and routinely screened simultaneously and in a non-destructive manner. Currently, flow cytometry is practically limited to 6-10 markers due to spectral emission overlap of the different fluorochromes used simultaneously. 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. This high-risk, high-reward R21 project has two Specific Aims: • Aim 1: To optimize device coding and processing for high throughput screening and real-time analysis. We will design and develop a unique NPS platform based on Barker codes that enable high- resolution detection even with low signal-to-noise ratios (SNRs). Barker codes are binary signals that are often used in radar and telecommunications to which NPS is analogous. • Aim 2: To incorporate sorting technology onto the NPS platform developed in Aim 1. We intend to realize the full potential of NPS and integrate a sorting technology to the NPS platform. The sorting technology will be based on mechanical pressure actuation to sort cells rapidly into phenotypic sub- populations for downstream analysis and/or culture. NPS development and proof-of-principle will be based on screening and sorting breast-cancer cell lines, MCF10A, MCF-7, MDA-MB-436, and MDA-MB-231-all of which have different malignancy and metastatic status-for markers EpCAM, CD44, CD24, CD29, CD49f, CD133, Axl, MUC1, EGFR, and ErbB2. By focusing on these markers, we would have an immediate impact in studies involving characterizing sub-populations of circulating tumor cells from patients with metastatic breast cancer. Thus, our proof-of-principle for a fully developed NPS has high impact. The proposed integrated multi-marker NPS and sorting technology proposed has potential for transformative impact in a number of fields ranging from fundamental life sciences research to point-of-care diagnostics. For example, flow cytometry is the cornerstone to diagnosis for many of the hematologic malignancies. With our screening/sorting technique, we could detect minimal residual disease and remission states. Beyond clinical diagnosis, our technology could be employed to characterize, for instance, changes in surface-marker expression during stem-cell differentiation in order to identify and isolate potentially important and rare sub-populations.

描述：该项目的总体目标是开发节点孔传感（NPS），这是一种创新的无标记微流体技术，使其在标记物数量方面超越流式细胞术，可以同时进行实际和常规筛选，并且目前，由于同时使用的不同荧光染料的光谱发射重叠测量细胞的通过时间，流式细胞术实际上仅限于 6-10 个标记。因为它与微流体通道中功能化的抗体相互作用（特异性或非特异性），该微流体通道已被节点分段。细胞表面受体和功能化抗体之间的特异性相互作用阻碍了细胞，导致更长的传输时间和随后对特定特定物质的测定。这个高风险、高回报的 R21 项目有两个具体目标： • 目标1：优化设备编码和处理以实现高通量筛选和实时分析。我们将设计和开发基于Barker 编码的独特NPS 平台，即使在低信噪比(SNR) 的情况下也能实现高分辨率检测。 ）巴克码是雷达和电信中常用的二进制信号，与 NPS 类似。 • 目标2：将分选技术整合到目标1 中开发的NPS 平台上。我们打算充分发挥NPS 的潜力，并将分选技术集成到NPS 平台中。分选技术将基于机械压力驱动来快速分选细胞。下游分析和/或培养的表型亚群将基于乳腺癌细胞系 MCF10A、MCF-7 的筛选和分类。 MDA-MB-436 和 MDA-MB-231 - 均具有不同的恶性和转移状态 - 对于标记物 EpCAM、CD44、CD24、CD29、CD49f、CD133、Axl、MUC1、EGFR 和 ErbB2 通过关注这些。标记物，我们将对涉及转移性乳腺癌患者循环肿瘤细胞亚群特征的研究产生直接影响。完全开发的 NPS 的原理验证具有很高的影响力，所提出的集成多标记 NPS 和分选技术具有在从基础生命科学研究到护理点诊断等多个领域产生变革性影响的潜力。例如，流式细胞术是诊断许多血液恶性肿瘤的基石，通过我们的筛查/分选技术，我们可以检测微小的残留疾病和缓解状态，除了临床诊断之外，我们的技术还可以用于表征疾病的变化。干细胞分化过程中的表面标记表达，以识别和分离潜在重要和稀有的亚群。