基于癌细胞表面电荷特性的循环肿瘤细胞高效捕获与无损释放纳米技术在临床诊断中的应用研究

Circulating tumor cells (CTC) detection holds great significance in terms of cancer early diagnoses, patients prognosis assessment, efficacy monitoring and personalized treatment. At present, most of the CTC detection technologies are mainly based on immune affinity or the characters of cancer cell sizes or dielectric property. However, these targeted properties are not specific to cancer cells. The numbers of CTC detected by the early methods were extremely low. We recently We recently reported that negative electrical charge is a specific cell surface property of all tested cancer cells. These findings were made by using a new nanotechnology we developed for measuring cell surface charges (Theranostics,2016,6(11):1887). Based on the above findings, we propose to design a new CTC detection technology in this study. Graphene-magnetic nanoparticles with high specific surface area will be used as detection nanoprobe. Then the surface of nanoprobe will be functionalized with poly(vinylamine-sodium acrylate), which have the property of charge reversible. By targeting the surface negative charge of cancer cells, we will perform the efficiently capturing and nondestructive release of CTC in three well-designed clinical tumor models of acute leukemia, late-stage lung cancer and bladder carcinoma in situ (CIS). In the model of leukemia, we will assess the real recovery rate of CTC. In the model of lung cancer, we will find whether accurate CTC enumeration reflect the changes in clinical status of the disease. In the bladder carcinoma model, we will determine the relationship between the appearance of CTC and metastasis by monitoring CTC during the course of disease.This proposal aims to develop a new CTC detection technology with the properties of high sensitivity, high accuracy, high efficiency and universality.

循环肿瘤细胞(CTC)检测对癌症早期诊断、患者预后评估、疗效监测及个性化治疗等都有重大意义。目前常用的CTC检测技术主要基于免疫亲和或利用癌细胞尺寸、介电等特征，然而这些靶向特征都不是癌细胞的独特性质，检测的CTC数量都非常少。我们最近利用纳米探针检测细胞表面电荷，发现癌细胞具有表面负电荷的普遍特性（Theranostics,2016,6(11):1887）。据此，本项目设计新型CTC检测技术。以高比表面积的石墨烯-磁性颗粒为探针，表面修饰电荷可反转的聚(乙烯胺-丙烯酸钠)，以癌细胞表面负电荷为靶标，在精心设计的三种临床肿瘤模型（急性白血病、晚期肺癌和膀胱原位癌）中进行CTC的高效捕获和无损释放。通过白血病模型评估CTC检测的真实回收率，在肺癌模型中探索CTC计数与疾病状态的关系，在膀胱癌模型中全程跟踪疾病从原发到转移全过程的CTC变化。发展一种灵敏、准确、高效和广谱的新型CTC检测技术。

循环肿瘤细胞(CTC)检测对癌症早期诊断、患者预后评估、疗效监测及个性化治疗等都有重大意义。本项目通过识别癌细胞功能的多功能纳米探针，并设计一套独创的CTC检测流程，实现对CTC非抗体依赖的快速、灵敏、特异、广谱、高效、无损检测。该新型CTC检测纳米技术适用于从白血病到实体癌的各种类型癌症，检测的CTC绝对数量比现有常用检测技术（免疫磁珠正向富集或负向富集法）高出2-3个数量级，最终富集到的CTC可以在镜下观察其形态，提高检测的准确度和特异性，全流程中无涉及昂贵的检测仪器设备。通过白血病模型评估CTC检测的真实回收率，在肺癌模型中探索CTC计数与疾病状态的关系，在膀胱癌模型中全程跟踪疾病从原发到转移全过程的CTC变化。克服了当前CTC检测技术灵敏度低、广谱性差、价格昂贵等瓶颈问题。解决目前循环肿瘤细胞检测领域技术瓶颈问题，引领癌症液体活检在临床实际应用的突破性进展。