Nielsen Helle H, Beck Hans C, Kristensen Lars P, Burton Mark, Csepany Tunde, Simo Magdolna, Dioszeghy Peter, Sejbaek Tobias, Grebing Manuela, Heegaard Niels H H, Illes Zsolt
Department of Neurology, Odense University Hospital, Odense, Denmark; Institute for Clinical Research, University of Southern Denmark, Odense, Denmark.
Center for Clinical Proteomics, Odense University Hospital, Odense, Denmark.
PLoS One. 2015 Oct 13;10(10):e0139659. doi: 10.1371/journal.pone.0139659. eCollection 2015.
Inflammatory demyelinating diseases of the CNS comprise a broad spectrum of diseases like neuromyelitis optica (NMO), NMO spectrum disorders (NMO-SD) and multiple sclerosis (MS). Despite clear classification criteria, differentiation can be difficult. We hypothesized that the urine proteome may differentiate NMO from MS.
The proteins in urine samples from anti-aquaporin 4 (AQP4) seropositive NMO/NMO-SD patients (n = 32), patients with MS (n = 46) and healthy subjects (HS, n = 31) were examined by quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) after trypsin digestion and iTRAQ labelling. Immunoglobulins (Ig) in the urine were validated by nephelometry in an independent cohort (n = 9-10 pr. groups).
The analysis identified a total of 1112 different proteins of which 333 were shared by all 109 subjects. Cluster analysis revealed differences in the urine proteome of NMO/NMO-SD compared to HS and MS. Principal component analysis also suggested that the NMO/NMO-SD proteome profile was useful for classification. Multivariate regression analysis revealed a 3-protein profile for the NMO/NMO-SD versus HS discrimination, a 6-protein profile for NMO/NMO-SD versus MS discrimination and an 11-protein profile for MS versus HS discrimination. All protein panels yielded highly significant ROC curves (AUC in all cases >0.85, p≤0.0002). Nephelometry confirmed the presence of increased Ig-light chains in the urine of patients with NMO/NMO-SD.
The urine proteome profile of patients with NMO/NMO-SD is different from MS and HS. This may reflect differences in the pathogenesis of NMO/NMO-SD versus MS and suggests that urine may be a potential source of biomarkers differentiating NMO/NMO-SD from MS.
中枢神经系统炎性脱髓鞘疾病包括多种疾病，如视神经脊髓炎（NMO）、视神经脊髓炎谱系障碍（NMO-SD）和多发性硬化症（MS）。尽管有明确的分类标准，但鉴别仍可能存在困难。我们推测尿液蛋白质组可能有助于区分NMO和MS。
对抗水通道蛋白4（AQP4）血清阳性的NMO/NMO-SD患者（n = 32）、MS患者（n = 46）和健康受试者（HS，n = 31）的尿液样本中的蛋白质进行胰蛋白酶消化和iTRAQ标记后，通过定量液相色谱-串联质谱（LC-MS/MS）进行检测。在一个独立队列（每组n = 9 - 10）中通过比浊法验证尿液中的免疫球蛋白（Ig）。
分析共鉴定出1112种不同蛋白质，其中109名受试者共有333种。聚类分析显示，与HS和MS相比，NMO/NMO-SD患者的尿液蛋白质组存在差异。主成分分析也表明NMO/NMO-SD蛋白质组图谱有助于分类。多变量回归分析显示，用于区分NMO/NMO-SD与HS的有3种蛋白质的图谱，区分NMO/NMO-SD与MS的有6种蛋白质的图谱，区分MS与HS的有11种蛋白质的图谱。所有蛋白质组均产生了高度显著的ROC曲线（所有情况下AUC>0.85，p≤0.0002）。比浊法证实NMO/NMO-SD患者尿液中Ig轻链增加。
NMO/NMO-SD患者的尿液蛋白质组图谱与MS和HS不同。这可能反映了NMO/NMO-SD与MS发病机制的差异，并表明尿液可能是区分NMO/NMO-SD与MS的生物标志物的潜在来源。
