单颗粒外泌体的表面蛋白特征表达谱分析新方法及其在癌症诊断中的应用研究

Exosomes, which contain a large amount of biological information derived from parental cells and play important roles in disease initiation and progression, are emerging biomarkers with great significance. However, exosome analysis at single particle level remains a huge challenge. In this project, we aim to develop a novel method for surface protein signature profiling of single exosome, and ultimately apply it to cancer diagnosis. Molecular probes are to be judiciously designed for specifically recognizing the potential protein biomarkers on exosome derived from different subtypes of liver cancer cells. The interactions between molecular probes and exosomes are to be investigated, and the mechanisms are to be elucidated as well. Novel assembly strategies are to be explored, so as to further conduct the construction of DNA nanostructure on the target exosome surface. Various parameters of flow cytometer are to be optimized. Under the optimized conditions, a novel method for surface protein signature profiling of single exosomes derived from different subtypes of liver cancer cells will be established. Finally, we devote to achieve discrimination of exosomes derived from different subtypes of cancer cells in clinical samples based on the differences on the expression levels of two and more surface biomarkers. This project will provide a novel and powerful method for the exosome-based fluid biopsy field. It holds great significance in the fields of early diagnostics, personalized treatment and precision medicine of cancerous diseases.

外泌体包含了大量来源于亲代细胞的生物信息，在疾病的发生和发展过程中具有重要的作用，因此是一种新型的重要生物标志物。然而，在单颗粒水平上对外泌体进行分析却存在着巨大的挑战。本项目拟开展单颗粒外泌体的表面蛋白特征表达谱分析新方法及其在癌症诊断中的应用研究。针对外泌体的特征表面蛋白设计分子探针以特异性识别不同亚型肝癌细胞外泌体，并研究探针与外泌体的相互作用、规律和机理；探索新的组装策略，在目标外泌体表面构建纳米结构；优化流式细胞仪参数，建立亚型癌细胞分泌的单颗粒外泌体的特征表达谱分析新方法；基于两种及两种以上表面特征蛋白的表达差异，实现不同亚型癌细胞外泌体的区分与鉴别。此项工作将为基于外泌体的液体活检领域提供一种十分有效的新方法，在癌症的早期诊断、个性化治疗以及精准医疗等领域具有重要的意义。