基于微阵列芯片检测的循环肿瘤细胞多靶标识别及单细胞分析新方法研究

The detection of circulating tumor cells (CTCs) is the he most potential technologies for liquid biopsy. However, recognition, isolation and characterization of CTCs have been challenging owing to their extremely low content, complex samples and dynamic nature. Herein, a system that consist of multi-target binding and trapping, multi-site recognition and detection as well as single cell gene analysis is to be developed aiming to solve the critical problems on CTCs analysis. Firstly, new affinity peptides will be designed and screened out towards the epithelial-mesenchymal transition process. Peptide functionalized nano materials with topological structures are to be developed for effective tumor cell isolation. Secondly, switchable peptide-fluorescent molecules are going to be developed to identify the molecular phenotypes of CTCs. Thirdly, one type of microarray chip is to be developed. On-line sorting and single cell trapping are going to be performed and multi-color fluorescent coding detection is going to be achieved on the chip. Finally, DNA primer library microarray is going to be constructed through combinatorial chemistry method. High-throughput single cell gene analysis is to be realized. The method proposed in this project provide new ideas and new techniques for tumor early diagnosis, molecular typing and prognosis evaluation.

循环肿瘤细胞（CTCs）检测是目前最具发展潜力的肿瘤液体活检技术。然而含量极低、样品复杂、动态变化等特点使得识别、分离以及鉴定CTCs成为难点，也成为生命分析领域的热点前沿。本项目瞄准CTCs分析的关键问题，提出了发展针对CTCs的多靶点结合捕获，多表位识别检测及单细胞基因分析体系。首先，聚焦肿瘤发展的上皮-间质转化过程，设计筛选亲和肽段，发展具有拓扑结构的新型多肽纳米材料高效捕获CTCs。其次，发展荧光“开关”型靶向多肽，实现CTCs的分子表型区分。进而，基于微阵列芯片，发展CTCs的在线分选和单细胞捕获，实现多色荧光编码的多靶标检测。最终，通过组合化学DNA引物文库微阵列，实现CTCs的高通量单细胞基因分析。项目提出的方法为肿瘤早期诊断、分子分型及预后评估等临床研究提供新的思路与新的技术。

本项目针对循环肿瘤细胞的精准分析和活体检测等方面的关键问题，发展了针对CTCs的多靶点结合捕获，多表位识别检测及多组学基因分析体系，产生了一系列科研成果。首先，聚焦肿瘤发展的上皮-间质转化过程，设计筛选亲和肽段，发展具有拓扑结构的新型多肽纳米材料高效捕获CTCs。其次，发展荧光“开关”型靶向多肽，实现CTCs的分子表型区分。进而，基于微阵列芯片，发展CTCs的在线分选和单细胞捕获，实现多色荧光编码的多靶标检测，实现CTCs的高通量单细胞基因分析。最终，实现了上皮间质转化过程中若干分子靶标的体内-体外一体化分子传感与检测。该项目为肿瘤早期诊断、分子分型及预后评估等临床研究提供新的思路与新的方法。