Identification of potential saliva and tear biomarkers in primary Sjögren's syndrome, utilising the extraction of extracellular vesicles and proteomics analysis.
Identification of potential saliva and tear biomarkers in primary Sjögren's syndrome, utilising the extraction of extracellular vesicles and proteomics analysis.
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利用细胞外囊泡和蛋白质组学分析的提取，在原发性Sjögren综合征中鉴定了潜在的唾液和撕裂生物标志物。
DOI：
10.1186/s13075-017-1228-x
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发表时间：
2017-01-25
影响因子：
4.9
通讯作者：
Jensen JL
中科院分区：
文献类型：
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作者：
Aqrawi LA;Galtung HK;Vestad B;Øvstebø R;Thiede B;Rusthen S;Young A;Guerreiro EM;Utheim TP;Chen X;Utheim ØA;Palm Ø;Jensen JL
在评估原发性干燥综合征（pSS）患者时，长期需要无创、更准确的诊断技术。纳入额外的诊断，包括筛选生物流体中的疾病特异性生物标志物是一个有前途的概念，需要进一步研究。在目前的研究中，我们的目的是探索pSS患者唾液和泪液中的新疾病生物标志物。对来自27名pSS患者和32名健康对照的刺激的全唾液和泪液进行液相色谱-质谱（LC-MS），并生成唾液和泪液蛋白质组生物标志物谱。还将LC-MS与尺寸排阻色谱结合以从两种流体中分离细胞外囊泡（EV）。对来自唾液和泪液的关节部分进行纳米颗粒跟踪分析，以确定EV的尺寸分布和浓度。通过使用磁珠免疫亲和捕获CD 9阳性EV进行进一步的EV表征，通过流式细胞术检测。使用Scaffold分析LC-MS数据的患者组和对照组之间的定量差异，并使用注释、可视化和集成发现数据库（大卫）进一步分析蛋白质的基因本体过度表达，以及使用检索相互作用基因/蛋白质的搜索工具进行蛋白质-蛋白质相互作用网络分析。在pSS的唾液中检测到参与先天免疫（LCN 2）、细胞信号传导（CALM）和创伤修复（GRN和CALML 5）的蛋白质的上调。唾液EV还显示出对先天免疫系统（SIRPA和LSP 1）和脂肪细胞分化（APMAP）的激活至关重要的生物标志物。泪液分析表明TNF-α信号传导（CPNE 1）和B细胞存活（PRDX 3）相关蛋白质的过表达。此外，中性粒细胞明胶酶相关的脂质运载蛋白上调唾液和泪液中的pSS。一致地，大卫分析证明了pSS患者唾液中的适应性免疫应答、唾液EV的细胞组分组装以及泪液中的代谢和蛋白质折叠的途径。来自pSS患者的唾液和泪液的LC-MS，单独和与尺寸排阻色谱法组合，允许筛选涵盖唾液和泪腺疾病靶器官的可能的新生物标志物。这种方法可以提供额外的诊断准确性pSS，也可能被应用于分期和监测疾病。本文的在线版本（doi：10.1186/s13075-017-1228-x）包含补充材料，可供授权用户使用。
There is a long-lasting need for non-invasive, more accurate diagnostic techniques when evaluating primary Sjögren’s syndrome (pSS) patients. Incorporation of additional diagnostics involving screening for disease-specific biomarkers in biological fluid is a promising concept that requires further investigation. In the current study we aimed to explore novel disease biomarkers in saliva and tears from pSS patients. Liquid chromatography-mass spectrometry (LC-MS) was performed on stimulated whole saliva and tears from 27 pSS patients and 32 healthy controls, and salivary and tear proteomic biomarker profiles were generated. LC-MS was also combined with size exclusion chromatography to isolate extracellular vesicles (EVs) from both fluids. Nanoparticle tracking analysis was conducted on joint fractions from the saliva and tears to determine size distribution and concentration of EVs. Further EV characterisation was performed by immunoaffinity capture of CD9-positive EVs using magnetic beads, detected by flow cytometry. The LC-MS data were analysed for quantitative differences between patient and control groups using Scaffold, and the proteins were further analysed using the Database for Annotation, Visualization and Integrated Discovery (DAVID), for gene ontology overrepresentation, and the Search Tool for the Retrieval of Interacting Genes/Proteins for protein-protein interaction network analysis. Upregulation of proteins involved in innate immunity (LCN2), cell signalling (CALM) and wound repair (GRN and CALML5) were detected in saliva in pSS. Saliva EVs also displayed biomarkers critical for activation of the innate immune system (SIRPA and LSP1) and adipocyte differentiation (APMAP). Tear analysis indicated overexpression of proteins involved in TNF-α signalling (CPNE1) and B cell survival (PRDX3). Moreover, neutrophil gelatinase-associated lipocalin was upregulated in saliva and tears in pSS. Consistently, DAVID analysis demonstrated pathways of the adaptive immune response in saliva, of cellular component assembly for saliva EVs, and of metabolism and protein folding in tears in pSS patients. LC-MS of saliva and tears from pSS patients, solely and in combination with size-exclusion chromatography allowed screening for possible novel biomarkers encompassing both salivary and lacrimal disease target organs. This approach could provide additional diagnostic accuracy in pSS, and could possibly also be applied for staging and monitoring the disease. The online version of this article (doi:10.1186/s13075-017-1228-x) contains supplementary material, which is available to authorized users.