多组学/深度裂解有机质谱流式技术及其在癌细胞巨胞饮监测中的应用

癌细胞巨胞饮作用是癌症化疗耐药的重要原因，有望成为癌症治疗的突破点。巨胞饮过程涉及细胞/囊泡内蛋白质和代谢物多组学物质动态转变、且存在明显的细胞异质性，其分子机制研究颇具挑战性。有机质谱流式作为一种新兴的单细胞分析模式，在多物质实时分析上具有极大潜力。但由于存在单细胞蛋白质检测困难、胞内物质检出有限等技术瓶颈，难以用于巨胞饮研究。本项目旨在发展多组学/深度裂解有机质谱流式分析平台，实现代表性的单个胰腺癌细胞巨胞饮过程中多组学的同时、实时监测。拟采用微/纳秒组合式脉冲高压作为喷雾电源实现离子化过程中的深度细胞裂解，从而提高对胞内、巨胞饮体内物质的检测覆盖度；设计并合成适用性强的乙二醇-吡啶盐-烷基硫醇系列质谱标签及相关探针，用于游离蛋白及膜蛋白的标记，实现单细胞中蛋白质和代谢物的同时监测。本项目的实施对于推动有机质谱流式分析技术的发展及其在癌细胞巨胞饮研究中的应用具有重要意义。

Macropinocytosis in cancer cells has been demonstrated to be crucial to chemotherapy resistance and promised to be a breakthrough in cancer treatment. However, the micropinocytosis process involves the dynamic changes of multi-omics including proteins and metabolites in cells/macropinosomes and presents large heterogeneity between individual cells, which makes the exploration of its molecular mechanism a big challenge. Organic mass cytometry is a promising technique for single cell analysis with the advantages of rich information and real-time analysis, but the insufficient sensitivity for single-cell proteins and the low coverage of intracellular compounds limit its application for single-cell micropinocytosis monitoring. The project aims to develop multi-omics/deep-lysing organic mass cytometry platform, and apply this platform for the monitoring of micropinocytosis-related multi-omic compounds inside representative single pancreatic cancer cells. For this purpose, combined micro/nano-second pulse voltage will be employed as spray power to deepen the cell lysis during ionization, ensuring the high information coverage of intracellular and intra-macropinosome targets. Furthermore, series of glycol-pyridinium-alkylthiol molecules will be prepared as versatile mass tags and further mass probes to label free proteins and membrane proteins, ensuring the simultaneous detection of proteins and metabolites during cytometric analysis. This project will play an important role in promoting the performance of organic mass cytometry technique and its application for studying micropinocytosis in cancer cells.