Proteomic and histopathological characterisation of sicca subjects and primary Sjogren's syndrome patients reveals promising tear, saliva and extracellular vesicle disease biomarkers
Proteomic and histopathological characterisation of sicca subjects and primary Sjogren's syndrome patients reveals promising tear, saliva and extracellular vesicle disease biomarkers
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DOI：
10.1186/s13075-019-1961-4
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发表时间：
2019-07-31
影响因子：
4.9
通讯作者：
Jensen, Janicke Liaaen
中科院分区：
文献类型：
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作者：
Aqrawi, Lara A.;Galtung, Hilde Kanli;Jensen, Janicke Liaaen
研究背景原发性干燥综合征（primarySjogren 'ssyndrome，pSS）的特征性表现是外分泌腺单核细胞浸润，产生Ro/SSA和La/SSB自身抗体，沿着口腔和眼部干燥。非SS干燥症受试者是一个与pSS相关的研究不足的群体，表现出类似的干燥症症状，唾液腺可能有轻度炎症迹象，但没有血清学检测到自身抗体产生。在这项研究中，我们调查的炎症表现在唾液腺组织，泪液和唾液的非SS受试者相比，pSS患者和健康individual.MethodsFifteen非SS，10 pSS和10名健康受试者被纳入分析。对唾液腺活检进行组织学评价。对泪液和刺激的全唾液进行液相色谱-质谱（LC-MS），并生成蛋白质组生物标志物谱。还将来自两种流体的细胞外囊泡（EV）分离和表征与LC-MS组合。使用Scaffold分析LC-MS数据以获得患者组和对照组之间的定量差异。数据库注释，可视化和集成发现（大卫）和功能丰富的分析工具（FunRich）被应用于functional analysis. ResultsHistological评价唾液腺活检显示，通过单核细胞浸润，纤维化和脂肪替代，在非SS受试者的炎症较轻的影响，相比，pSS患者。虽然在非SS组中未受影响，但在pSS患者的泪液中检测到促炎途径和参与泛素化（LMO 7和HUWE 1）和B细胞分化（TPD 52）的蛋白的上调。此外，在来自pSS患者泪液的EV中进一步鉴定了调节细胞先天性和适应性免疫途径的蛋白STOM、ANXA 4和ANXA 1的过表达。最后，全唾液和EV分离的pSS患者的全唾液表达的蛋白质至关重要的先天性MHC I类细胞调节（NGAL）和T细胞活化（CD 44）。ConclusionsNon-SS干燥症受试者可能会显示轻度炎症的影响，在他们的腺体组织，而他们的蛋白质谱是惊人的更相似的健康对照组比pSS患者。因此，所鉴定的泪液和唾液生物标志物可以作为潜在的非侵入性诊断工具来实施，其可以在评估非SS受试者和pSS患者以及监测疾病进展时帮助提高诊断准确性。
BackgroundMononuclear cell infiltration of exocrine glands, production of Ro/SSA and La/SSB autoantibodies, along with oral and ocular dryness, are characteristic features of primary Sjogren's syndrome (pSS). Non-SS sicca subjects, an underexplored group in relation to pSS, display similar sicca symptoms, with possible mild signs of inflammation in their salivary glands, yet with no serological detection of autoantibody production. In this study, we investigated inflammatory manifestations in the salivary gland tissue, tear fluid and saliva of non-SS subjects, as compared to pSS patients and healthy individuals.MethodsFifteen non-SS, 10 pSS and 10 healthy subjects were included in the analyses. Histological evaluation of salivary gland biopsies was performed. Liquid chromatography-mass spectrometry (LC-MS) was conducted on tear fluid and stimulated whole saliva, and proteomic biomarker profiles were generated. Extracellular vesicle (EVs) isolation and characterisation from both fluids were also combined with LC-MS. The LC-MS data were analysed for quantitative differences between patient and control groups using Scaffold. Database for Annotation, Visualization and Integrated Discovery (DAVID) and Functional Enrichment Analysis Tool (FunRich) were applied for functional analyses.ResultsHistopathological evaluation of salivary gland biopsies showed implications of milder inflammation in non-SS subjects through mononuclear cell infiltration, fibrosis and fatty replacement, as compared to pSS patients. Although unaffected in the non-SS group, upregulation of proinflammatory pathways and proteins involved in ubiquitination (LMO7 and HUWE1) and B cell differentiation (TPD52) were detected in tear fluid of pSS patients. Moreover, overexpression of proteins STOM, ANXA4 and ANXA1, regulating cellular innate and adaptive immunological pathways, were further identified in EVs from tear fluid of pSS patients. Finally, whole saliva and EVs isolated from whole saliva of pSS patients expressed proteins vital for innate MHC class I cellular regulation (NGAL) and T cell activation (CD44).ConclusionsNon-SS sicca subjects may show implications of mild inflammation in their glandular tissue, while their protein profile was strikingly more similar to healthy controls than to pSS patients. Hence, the tear and salivary biomarkers identified could be implemented as potential non-invasive diagnostic tools that may aid in increasing diagnostic accuracy when evaluating non-SS subjects and pSS patients and monitoring disease progression.