High throughput tear proteomics with data independent acquisition enables biomarker discovery in allergic conditions

The search for pathological biomarkers in biological fluids that can provide valuable insight into an individual’s health status, is a relevant area of research for multiple pathologies. Currently, the use of proteomics for the identification of differences in protein expression profiles between samples from healthy subjects and patients, has emerged as a powerful strategy to improve the current diagnosis of various pathologies or propose novel therapeutic approaches. Among the biological fluids from which new pathological biomarkers can be identified, tear secretion is highly attractive, since it can be collected non-invasively and could better concentrate proteins that sensitively reflect allergic responses, owing to their exposure to environmental factors and its connection to the respiratory system. Despite its potential, tear fluid remains underexplored, offering significant research opportunities. In this study, we collected human tear samples using the Shirmer Test from healthy and allergic individuals. Our optimized workflow, combining sample preparation and high-throughput proteomics using the data-independent acquisition (DIA) strategy, identified 2,542 proteins and enabled the successful differentiation of the two groups. We identified 99 differentially expressed proteins. Our results show the feasibility of protein analysis in human tear samples, highlighting tears as a highly sensitive fluid for detecting health conditions.

探索能够为个体健康状态提供宝贵参考信息的生物体液中的病理生物标志物，是针对多种疾病的重要研究方向。当前，利用蛋白质组学（proteomics）分析健康受试者与患者样本间的蛋白质表达谱差异，已成为优化多种疾病现有诊断方案、或提出全新治疗策略的有力手段。在可用于发掘新型病理生物标志物的各类生物体液中，泪液极具研究价值：其采集方式无创，且因暴露于环境因素且与呼吸系统相连，能够更好地富集可灵敏反映过敏反应的蛋白质。尽管具备上述潜力，泪液仍未得到充分探索，蕴含着可观的研究空间。本研究采用施墨试验（Shirmer Test）采集了健康个体与过敏患者的人泪液样本。本研究优化的实验流程整合了样本制备与采用数据非依赖采集（DIA）策略的高通量蛋白质组学技术，共鉴定出2542种蛋白质，并成功实现了两组样本的区分。本研究共鉴定出99种差异表达蛋白质。研究结果证实了人泪液样本蛋白质分析的可行性，凸显出泪液作为可灵敏检测健康状态的体液的应用价值。