基于胸水中微颗粒代谢组检测技术鉴别良、恶性胸腔积液的新方法研究

Pleural effusions are the common disease of respiratory system. Accurate and rapid diagnosis of begnign pleural effudions (BPE) and malignant pleural effudions (MPE) is the key to efficient treatment of pleural effusions, but it is lack of sensitive and specific early diagnostic method. The intercellular messenger of microparticles are considered to the ideal diagnostic biomarkers carriers, and the metabolomics study can provide a fast and sensitive discovery of the disease early alteration, in the previous study we found the levels of most metabolites in microparticles were significantly differential between BPE and MPE, which illustrates the detection of metabolome in microparticles may provide a new method for the diagnosis of BPE and MPE. However, current studies on metabolome in microparticles are very limited, so it is very necessary to develop new high-sensitive and high-specific metabolome detection methods of microparticles. . Based on these, we intend to do further study as following: 1.development of high-sensitive and high-specific metabolome detection methods of microparticles by pseudo-targeted metabolomics technique. 2. Investigation the metabolic alteration in microparticles between BPE and MPE by these methods, and screening novel biomarkers for the diagnosis of BPE and MPE by the pathway enrichment analysis and correlation analysis between clinical data and differential metabolites. 3. Establishment a novel diagnostic model based on these identified biomarkers, whose diagnostic sensitivity and specificity are further evaluated by other clinical samples. This study will develop a high-sensitive and high-specific metabolome detection for microparticles, and provide a novel method for the early diagnosis of diseases.

胸腔积液是一种呼吸系统常见疾病，准确快速诊断其良、恶性是有效治疗胸腔积液的关键，目前临床缺乏敏感特异性的早期诊断指标。研究证实细胞间信使-微颗粒是理想的疾病诊断标志物载体，代谢组学研究可快速灵敏地发现疾病的早期变化，且我们前期研究发现良、恶性胸水中微颗粒代谢物分子水平有显著性差异，表明胸水中微颗粒代谢组检测可为良、恶性胸腔积液鉴别提供新方法。目前系统检测微颗粒代谢组的方法较为缺乏，因此有必要发展新型灵敏特异性强的检测方法。. 基于此，我们拟进一步研究：1.利用LC-MS拟靶标代谢组学技术建立高灵敏度、高特异性的微颗粒代谢组检测方法；2.系统比较良、恶性胸水中微颗粒代谢组，结合代谢通路富集分析及临床数据相关性分析，筛选良、恶性胸腔积液诊断标志物；3.建立诊断模型，并开展临床验证，评估其诊断效果。本研究将创建一种高灵敏、高特异性的微颗粒代谢组检测方法，并为疾病的早期诊断研究提供新途径。

胸腔积液是全球常见的呼吸系统疾病，但其良恶性诊断仍然具有挑战性，亟需发展新型稳定可靠的早期诊断方法。细胞外微颗粒(EVs)已被证实为疾病生物标志物有希望的来源，但其代谢物组成及其在胸腔积液诊断中的作用尚不清楚。本项目建立了基于LC-MS技术的高灵敏度、高特异性的微颗粒代谢组检测方法；同时，结合差异超速离心法分离提取胸水中的两个EV亚群，即大细胞外微颗粒(lEVs)和小细胞外微颗粒(sEVs)，系统比较了在结核和恶性胸腔积液组间两者的代谢组特征，并筛选了诊断标志物及建立了诊断模型，评估其诊断效果。.研究结果显示，胸水EVs中共检测到579种代谢产物，包括氨基酸、酰基肉碱、有机酸、类固醇、酰胺和各种脂类物质。其中，lEVs和sEVs的代谢谱有重叠和差异，且与胸水的代谢谱有显著差异。同时，良恶性胸腔积液中不同亚型的EVs表现出独特的代谢富集。此外，相比较sEVs的结果，lEVs在结核组(TPE)和恶性组(MPE)之间表现出更显著和更大的代谢变化，其所涉及的差异代谢物与临床参数的相关性更密切。最后，结合系统的发现和验证工作流，确定了四种候选生物标志物，包括苯丙氨酸、亮氨酸、磷脂酰胆碱35:0和鞘磷脂44:3。相比较传统临床标志物，该组合标志物在鉴别TPE和MPE具有较高的灵敏度和特异性，特别是针对误诊或漏诊的患者，例如鉴别两组的受试者工作特征曲线（AUC）下的面积均> 0.95。.综上所述，本研究对胸水中的EVs进行了全面的代谢谱分析，并从其亚型lEVs和sEVs的代谢产物水平进一步探索了结核和恶性胸腔积液的代谢重编程，为深入了解胸腔积液的机制提供了依据，并为TPE和MPE的诊断提供了新途径。