基于线粒体靶向AIE探针的循环肿瘤细胞检测新技术及其机制研究

Circulating tumor cell (CTC) can provide important information for cancer management ranging from early diagnosis, dynamic monitoring, resistance assessing, therapy guiding, and recurrence monitoring non-invasively. However, the clinical applications and characterization of CTC on single cell level are still limited, largely attributed to the lack of ideal markers. In our previous work, TPN, a mitochondria-targeting fluorescent probe with aggregation-induced emission (AIE) characteristics was found to exhibit low toxicity, strong fluorescence, and excellent photostability. Furthermore, TPN can rapidly accumulate in mitochondria of live cancer cells with much higher fluorescence than that in white blood cells. According to these observations, we propose that TPN may become a new tool for live CTC identification based on mitochondrial differences between benign and malignant cells. This project aims to investigate and optimize the optimal conditions for tumor cells identification by TPN, followed by evaluating its clinical performance compared with traditional protein markers in CTC detection using whole blood samples. Moreover, the mechanism of TPN in tumor cells identification will also be investigated focusing on the mitochondrial biology. And we expect to obtain theoretical and experimental evidence from this research to develop a rapid, simple and non-invasive assay for circulating tumor cell detection.

循环肿瘤细胞（CTC）可以无创的方式提供肿瘤早期诊断、动态观察、疗效判断、用药指导以及复发监测等重要信息，然而CTC的临床应用与单细胞分析仍受限，其中一个重要原因是CTC的鉴定仍缺乏理想的标志物。在前期工作中，我们发现一种具有聚集诱导发光（AIE）效应的新型荧光分子TPN，其展现出低毒性、高荧光强度、高光稳定性以及细胞线粒体靶向的独特性质，可快速选择性聚集于肿瘤细胞线粒体中，使其呈现出比白细胞更高的荧光强度。据此，我们提出利用TPN有望发展一种基于线粒体差异的活CTC鉴定新方法。本项目拟进一步考察并优化TPN鉴定肿瘤细胞的最佳应用条件，并通过对实际临床全血标本CTC的检测以及与传统蛋白标志物的CTC鉴定方法进行对比，系统评估该方法的临床检测效能，并从线粒体数量、膜电位与TPN荧光信号关系方面深入探索作用机制，为建立一种快速、简便、无损的新型活循环肿瘤细胞检测技术提供理论与实验依据。

项目主要开发基于线粒体靶向新型荧光探针进行循环肿瘤细胞无损检测的新方法及临床研究。从肿瘤的旺盛的生命活动特征入手，基于肿瘤细胞的线粒体数量增多，同时具有更高的线粒体膜电位，使得具有线粒体靶向的新型聚集诱导发光生物探针TPN在肿瘤细胞中有更高的荧光强度。通过肿瘤细胞与白细胞荧光强度的差异，实现肿瘤细胞的鉴别。同时，TPN分子是水溶性的小分子探针，具有良好的生物相容性，突破传统鉴定技术需要对细胞进行固定、穿透等步骤，能无损的对细胞进行识别，可支持对肿瘤细胞的单细胞测序及后续基因信息的读取，有望促进CTC的临床应用，并为肿瘤用药指导、复发监测等精准医学需求提供新的窗口。本项目共发表SCI论文5篇（其中5篇IF＞10，总IF＞70）。获广东省科技进步一等奖1项、二等奖1项。协助培养博士研究生、硕士研究生各1名。