Institute of Food Safety, Chinese Academy of Inspection and Quarantine, Beijing 100176, China.
Key Laboratory of Food Quality and Safety for State Market Regulation, School of Pharmacy, China Medical University, Shenyang 110122, China.
Anal Chem. 2024 Aug 6;96(31):12817-12826. doi: 10.1021/acs.analchem.4c02332. Epub 2024 Jul 25.
Single-cell analysis by mass spectrometry (MS) is emerging as a powerful tool that not only contributes to cellular heterogeneity but also offers an unprecedented opportunity to predict pathology onset and facilitates novel biomarker discovery. However, the development of single-cell MS analysis techniques with a focus on sample extraction, separation, and ionization methods for volume-limited samples and complexity of cellular samples are still a big challenge. In this study, we present a high-throughput approach to inkjet drop on demand printing single-cell MS for rapid screening of biomarkers of polycyclic aromatic hydrocarbon (PAH) exposure at the KYSE-150 cell, aiming to elucidate the pathogenesis of PAH-induced esophageal cancer. With an analytical bulk KYSE-150 cell throughput of up to 51 cells per minute, the method provides a new opportunity for simultaneous single-cell analysis of multiple biomarkers. We screened 930 characteristic ions from 3,683 detected peak signals and identified 91 distinctive molecules that exhibited significant differences under various concentrations of PAH exposure. These molecules have potential as clinical diagnostic biomarkers. Additionally, the current study identifies specific biomarkers that behave completely opposite in single-cell and multicell lipidomics as the concentration of PAH changes. These biomarkers potentially subdivide KYSE-150 cells into PAH-sensitive and PAH-insensitive types, providing a basis for revealing PAH toxicity and disease pathogenesis from the heterogeneity of cellular metabolism.
单细胞分析的质谱分析（MS）正在成为一种强大的工具，不仅有助于细胞异质性，而且还提供了一个前所未有的机会来预测病理学发病，并促进新的生物标志物的发现。然而，开发专注于样本提取、分离和离子化方法的单细胞 MS 分析技术，以适应有限体积的样本和细胞样本的复杂性，仍然是一个巨大的挑战。在这项研究中，我们提出了一种高通量的按需喷墨单细胞 MS 方法，用于快速筛选多环芳烃（PAH）暴露的生物标志物，目的是阐明 PAH 诱导的食管癌的发病机制。该方法的分析批量 KYSE-150 细胞的通量高达每分钟 51 个细胞，为同时分析多个生物标志物的单细胞提供了新的机会。我们从 3683 个检测到的峰信号中筛选出 930 个特征离子，并鉴定出 91 个在不同浓度的 PAH 暴露下表现出显著差异的独特分子。这些分子具有作为临床诊断生物标志物的潜力。此外，本研究确定了在单细胞和多细胞脂质组学中，随着 PAH 浓度的变化，表现出完全相反行为的特定生物标志物。这些生物标志物可能将 KYSE-150 细胞细分为对 PAH 敏感和不敏感的类型，为从细胞代谢的异质性中揭示 PAH 毒性和疾病发病机制提供了基础。
