基于tmTNF-α mAb磁分离微流控技术的乳腺癌转移超早期预警研究

项目名称： 基于tmTNF-α mAb磁分离微流控技术的乳腺癌转移超早期预警研究

项目编号： No.81472033

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 喻明霞

作者单位： 武汉大学

项目金额： 72万元

中文摘要： 循环肿瘤细胞（CTCs）是监测肿瘤复发和早期转移预警的良好指标。由于肿瘤的异质性、EMT、超早期隐匿性转移、癌前炎性改变等因素, 使Cellsearch 检测CTCs临床应用受限。新近研究表明：TNF-α 协同其他分子在乳腺癌的成瘤和转移中发挥作重要用，成为超早期预警和干预的分子靶点。本课题组前期证实：1）TNF-α在多种肿瘤中高表达，而正常组织和癌旁鲜见；2）在不同类型乳腺癌和癌前各级增生中表达并与组织学分级正相关；3）自主研发的tmTNF-α mAb 能高效抑制乳腺癌生长和转移。本项目拟以tmTNF-α mAb 靶向识别、磁性微流控芯片细胞捕获和量子点荧光标记成像并进行光谱分析，建立一种磁性微流控芯片高灵敏、高通量、高特异的检测CTCs的新方法并进行临床验证，捕获后的细胞鉴定分子表征。为开发抗体的多元磁控技术提供佐证，为分子表征谱的鉴定以及实时CTCs的液体活检提供实验依据。

中文关键词： 微流控芯片；量子点；循环乳腺癌细胞；多元分析；单克隆抗体

英文摘要： Recent studies indicate that micrometastasis emerges in early phase of breast cancer, then detection of circulating tumor cells (CTCs) becomes a good indicator for monitoring relapse and early warning tumor metastasis. As a result of the heterogeneity of tumor, EMT, ultra-early occult metastasis, precancerous inflammatory lesion and failure to specifically distinguish normal-like CTCs, the application of Cellsearch assay was restricted, which was the only method approved by FDA. Researches showed that the development of breast cancer was closely related with inflammatory process, and that TNF-α, cooperating with other molecules, acted as an important role in the process of tumorigenesis and metastasis. Hence, TNF-α became a molecular target for ultra-early warn and intervention. Our research group turned out in previous works: 1) TNF-α was highly expressed in many tumors, but rarely in normal and peritumoral tissues; 2) It was also expressed in different subtypes of breast cancer and all grades of breast hyperplasia, which was positive correlated with histological grades; 3) The tmTNF-α monoclonal antibody that we developed independently could efficiently suppress breast cancer growth and metastasis. So, in this project, by means of a combination of triple steps-targeted recognition by tmTNF-α mAb, specific capture by magnetic microfluidic chip and precisely labeling by quantum dots for sensitive imaging and spectrum analysis, we plan to establish a new diagnostic approach for detecting CTCs with High sensitivity, high-throughput, and high specificity on the surface of the chip, and then conduct clinical validation. The cells we captured will be cultured in vitro for the further identity of their molecular expression profiling. As a result, our work will provide strong support for developing antibodies-based multiplex magnetic microfluidic, as well as provide experimental evidence for identity of molecular expression profiling and being liquid biopsy of real time CTCs in vivo.

英文关键词： microfluidics;QDs;Cirulating BC;multiplex assay;monoclonal antibody