Reich Michael, Dacheva Ivanka, Nobl Matthias, Siwy Justyna, Schanstra Joost P, Mullen William, Koch Frank H J, Kopitz Jürgen, Kretz Florian T A, Auffarth Gerd U, Koss Michael J
Eye Center, Albert-Ludwigs-University Freiburg, Freiburg, Germany.
Department of Ophthalmology, University of Heidelberg, Heidelberg, Germany.
PLoS One. 2016 Jun 30;11(6):e0158001. doi: 10.1371/journal.pone.0158001. eCollection 2016.
To analyze the protein profile of human vitreous of untreated patients with retinal vein occlusion (RVO).
Sixty-eight vitreous humor (VH) samples (44 from patients with treatment naïve RVO, 24 controls with idiopathic floaters) were analyzed in this clinical-experimental study using capillary electrophoresis coupled to mass spectrometer and tandem mass spectrometry. To define potential candidate protein markers of RVO, proteomic analysis was performed on RVO patients (n = 30) and compared with controls (n = 16). To determine validity of potential biomarker candidates in RVO, receiver operating characteristic (ROC) was performed by using proteome data of independent RVO (n = 14) and control samples (n = 8).
Ninety-four different proteins (736 tryptic peptides) could be identified. Sixteen proteins were found to be significant when comparing RVO and control samples (P = 1.43E-05 to 4.48E-02). Five proteins (Clusterin, Complement C3, Ig lambda-like polypeptide 5 (IGLL5), Opticin and Vitronectin), remained significant after using correction for multiple testing. These five proteins were also detected significant when comparing subgroups of RVO (central RVO, hemi-central RVO, branch RVO) to controls. Using independent samples ROC-Area under the curve was determined proving the validity of the results: Clusterin 0.884, Complement C3 0.955, IGLL5 1.000, Opticin 0.741, Vitronectin 0.786. In addition, validation through ELISA measurements was performed.
The results of the study reveal that the proteomic composition of VH differed significantly between the patients with RVO and the controls. The proteins identified may serve as potential biomarkers for pathogenesis induced by RVO.
分析未经治疗的视网膜静脉阻塞（RVO）患者玻璃体液的蛋白质谱。
在这项临床实验研究中，使用毛细管电泳结合质谱仪和串联质谱法对68份玻璃体液（VH）样本（44份来自初治RVO患者，24份来自特发性飞蚊症对照）进行分析。为了确定RVO的潜在候选蛋白质标志物，对RVO患者（n = 30）进行蛋白质组分析，并与对照组（n = 16）进行比较。为了确定RVO中潜在生物标志物候选物的有效性，使用独立RVO（n = 14）和对照样本（n = 8）的蛋白质组数据进行受试者操作特征（ROC）分析。
可鉴定出94种不同蛋白质（736个胰蛋白酶肽段）。比较RVO和对照样本时，发现16种蛋白质有显著差异（P = 1.43E - 05至4.48E - 02）。经过多重检验校正后，5种蛋白质（簇集蛋白、补体C3、免疫球蛋白λ样多肽5（IGLL5）、视蛋白和玻连蛋白）仍具有显著性。比较RVO亚组（中央RVO、半中央RVO、分支RVO）与对照组时，这5种蛋白质也有显著差异。使用独立样本确定曲线下面积（ROC - Area），证明结果的有效性：簇集蛋白0.884，补体C3 0.955，IGLL5 1.000，视蛋白0.741，玻连蛋白0.786。此外，还通过酶联免疫吸附测定（ELISA）测量进行了验证。
研究结果表明RVO患者与对照组的玻璃体液蛋白质组组成存在显著差异。鉴定出的蛋白质可能作为RVO诱导发病机制的潜在生物标志物。
