Integrated Microdevice to Capture and Detect Circulating Tumor Cells

描述（由申请人提供）：转移可能是决定癌症患者结局的最重要事件。 骨髓中隐匿性转移的检测虽然已知具有临床重要性，但尚未成为常规临床实践。 这是由于目前收集和检测这些物质的方法存在技术困难和成本问题。 血液中循环肿瘤细胞（CTC）的检测不如骨髓中的敏感，并且与骨髓中肿瘤细胞的检测存在相同的技术障碍，但是提供了侵入性更小并且患者依从性更好的明显优势。 因此，以微创和用户友好的方式敏感地检测肿瘤的最早转移扩散将对癌症患者的临床管理产生重大影响。 目前可用的四氯化碳捕获和鉴定方法面临重大障碍，包括程序步骤多、大量人为干预、成本极高，而且重要的是，检测方法缺乏可靠性和标准化。 我们已经证明了使用基于聚对二甲苯的生物膜进行基于大小的肿瘤细胞捕获的潜力。 我们建议将其开发成一种微芯片设备，用于处理血液，最终处理骨髓和其他液体，如胸腔积液或腹水。 这种微设备与微流体技术相结合，有可能彻底改变肿瘤细胞捕获和识别的方法。 此外，我们建议开发用于捕获的细胞的芯片上表征的方法。 首先，在R21阶段，我们将使用模型系统开发和优化捕获设备，以分离和分子表征混合在血液中的培养癌细胞，然后进行试点研究，检查45名患有乳腺癌，前列腺癌或膀胱癌转移性疾病的实际癌症患者的血液。 在R33阶段，我们将扩展microdevice的应用，以评估来自相同三种恶性肿瘤的约310名患者样本，我们还将使用量子点评估CTC的分子特征，以了解这些罕见细胞的生物学特征（例如存在可能更恶性的推定干细胞亚群）。 完成后，该项目的研究将开发一种具有成本效益的芯片系统，用于捕获，识别和表征CTC，易于在临床环境中使用。

DESCRIPTION (provided by applicant): Metastasis is probably the most important event for determining outcome in cancer patients. The detection of occult metastases in the bone marrow, while known to be clinically important, has not become routine clinical practice. This is due to the technical difficulties and costs involved in the current methods for their collection and detection. Detection of circulating tumor cells (CTC) in the blood is less sensitive than in bone marrow and suffers from the same technical barriers as the detection of tumor cells in the bone marrow, but offers the distinct advantage of being less invasive and better for patient compliance. Therefore, sensitive detection of earliest metastatic spread of tumor in a minimally invasive and user-friendly manner will have a great impact on the clinical management of cancer patients. The currently available methodologies for CTC capture and identification face significant barriers including multiple procedural steps, substantial human intervention, extremely high cost, and importantly, lack of reliability and standardization for the detection methods. We have demonstrated the potential for sized-based tumor cell capture using a parylene-based micropore membrane. We propose to develop this into a microchip device for processing blood, and eventually bone marrow and other fluids like pleural effusions or ascites. This microdevice, coupled with microfluidics, has the potential to revolutionize the approach to tumor cell capture and identification. Further, we propose to develop methods for on-chip characterization of the captured cells. First, in R21 Phase, we will develop and optimize the capture device using a model system to isolate and molecularly characterize cultured cancer cells admixed in blood, followed by a pilot study to examine blood from 45 actual cancer patients with metastatic disease for breast, prostate or bladder cancer. In R33 Phase, we will extend the application of microdevice to assess about 310 patient samples from the same three malignancies, and we will also assess the molecular characteristics of the CTC using the Quantum Dots to understand the biological features of these otherwise rare cells (such as existence of putative stem cell sub-population which may be more malignant). At completion, studies in this project will develop a cost effective on-chip system for capture, identification, and characterization of CTC, easily usable in the clinical setting.