Molecular Detection and Profiling of Circulating Tumor Cells

描述(由申请人提供)：拟议研究的长期目标是开发一种新型的传感器设备平台，能够检测和分析临床样本中的循环肿瘤细胞(CTCs)。CTCs是一种罕见的癌细胞，它从原发肿瘤部位逃逸并进入血流。在大多数转移性疾病患者和局部癌症患者的外周血中都发现了它们，但水平极低(大约每毫升血液中有1-100个细胞)。我们的假设是，这些外周细胞可以用于分子分析，可以无创性地为预后评估、抗癌药物开发以及最终基于CTCs的个性化癌症治疗提供有价值的信息。到目前为止，缺乏对这些细胞进行分子表征所需的技术，因为现有的临床技术过于不敏感。在之前的研究中，我们的实验室开发了一种原型微型核磁共振(p-核磁共振)传感器芯片，该芯片已被用于检测特定标记了超顺磁性纳米颗粒的癌细胞。这项技术被称为诊断磁共振(DMR)，在本提案1中，试图显著推进DMR技术，以便将其应用于临床用于CTCs的分子检测和图谱分析。为了实现这一目标，DMR检测的灵敏度和特异性将通过以下目标大幅提高：目标1.广泛优化用于使用针对细胞内和细胞外分子生物标记物的磁性纳米颗粒标记肿瘤细胞的程序，以及目标2.开发一种集成的微流体组件，根据大小将CTCs从血液中的其他细胞中分离出来，并将它们直接输送到p-核磁共振传感器。这项工作代表了一项多学科的努力，呼吁纳米技术、分子靶向、磁共振成像/传感和微流体领域的组件开发一种能够询问临床CTC样本的新型设备平台，该平台可以实现对肿瘤样本的实时分子分析，并在分子靶向治疗不断涌现的时代促进合理的治疗决策。这项研究在研究从血液样本中获取的人类癌细胞方面具有广泛的应用，因此可以非侵入性地获得信息，以评估患者的预后，设计治疗策略，并监测治疗反应。

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to develop a novel sensor device platform that is capable of detecting and molecularly profiling circulating tumor cells (CTCs) in clinical samples. CTCs are rare cancer cells that have escaped from a primary tumor site and entered the blood stream. They have been identified in the peripheral blood of most patients with metastatic disease as well as in patients with localized cancers, but at exceedingly low levels (approximately 1-100 cells per ml of blood). It is our hypothesis that these peripheral cells can be used for molecular analysis, which could non-invasively provide valuable information for use in prognosis evaluation, anticancer-drug development, and ultimately, personalized cancer therapy based on CTCs. To date, the technology required to molecularly characterize these cells is lacking because existing clinical technologies are too insensitive. In prior research, our lab developed a prototype micro- NMR (p-NMR) sensor chip that has been used to detect cancer cells that are specifically tagged with superparamagnetic nanoparticles. This technique has been termed diagnostic magnetic resonance (DMR), In this proposal 1 seek to significantly advance DMR technology so that it may be applied in clinical use for molecular detection and profiling of CTCs. To achieve this goal, DMR detection sensitivity and specificity will be dramatically increased through the following aims: Aim 1. Extensively optimize the procedures used to tag tumor cells with magnetic nanoparticles targeted to intracellular and extracellular molecular biomarkers, and Aim 2. Develop an integrated microfluidics component that isolates CTCs from other cells in the blood based on size and delivers them directly to the p-NMR sensor. This work represents a multidisciplinary effort that calls on components from the fields of nanotechnology, molecular targeting, magnetic resonance imaging/sensing, and microfluidics to develop a novel device platform capable of interrogating clinical CTC samples, which could enable real-time molecular analysis of tumor samples and facilitate rational treatment decisions in an era where molecularly targeted therapies are emerging. The proposed research has broad applications for studying human cancer cells that are obtained from blood samples, Information could thus be obtained non-invasively to evaluate patient prognosis, devise treatment strategies, and monitor therapeutic response.