Abstract：
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.
摘要：
通过质谱（MS）的单细胞分析正在成为一种强大的工具，不仅有助于细胞异质性，而且还提供了前所未有的机会来预测病理发作并促进新的生物标志物发现。然而,以样品提取为重点的单细胞质谱分析技术的发展，分离,和电离方法的体积有限的样品和细胞样品的复杂性仍然是一个很大的挑战。在这项研究中,我们提出了一种高通量的方法，以按需喷墨打印单细胞MS，用于在KYSE-150细胞中快速筛选多环芳烃（PAH）暴露的生物标志物，旨在阐明PAH诱导的食管癌的发病机制。分析批量KYSE-150细胞吞吐量高达每分钟51个细胞，该方法为同时进行多种生物标志物的单细胞分析提供了新的机会.我们从3,683个检测到的峰信号中筛选了930个特征离子，并鉴定了91个独特的分子，这些分子在各种浓度的PAH暴露下表现出显着差异。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与PAH浓度变化完全相反的特定生物标志物.这些生物标志物可能将KYSE-150细胞细分为PAH敏感型和PAH不敏感型。从细胞代谢的异质性为揭示PAH的毒性和疾病的发病机制提供了依据。
