Wasinger Valerie C, Yau Yunki, Duo Xizi, Zeng Ming, Campbell Beth, Shin Sean, Luber Raphael, Redmond Diane, Leong Rupert W L
From the ‡Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical centre, The University of New South Wales, Australia; §School of Medical Sciences, The University of New South Wales, Sydney, NSW, Australia;
From the ‡Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical centre, The University of New South Wales, Australia; ¶Gastroenterology Department, Concord Repatriation General Hospital, Hospital Rd, Concord, NSW, Australia;
Mol Cell Proteomics. 2016 Jan;15(1):256-65. doi: 10.1074/mcp.M115.055095. Epub 2015 Nov 3.
Breakdown of the protective gut barrier releases effector molecules and degradation products into the blood stream making serum and plasma ideal as a diagnostic medium. The enriched low mass proteome is unexplored as a source of differentiators for diagnosing and monitoring inflammatory bowel disease (IBD) activity, that is less invasive than colonoscopy. Differences in the enriched low mass plasma proteome (<25 kDa) were assessed by label-free quantitative mass-spectrometry. A panel of marker candidates were progressed to validation phase and "Tier-2" FDA-level validated quantitative assay. Proteins important in maintaining gut barrier function and homeostasis at the epithelial interface have been quantitated by multiple reaction monitoring in plasma and serum including both inflammatory; rheumatoid arthritis controls, and non-inflammatory healthy controls; ulcerative colitis (UC), and Crohn's disease (CD) patients. Detection by immunoblot confirmed presence at the protein level in plasma. Correlation analysis and receiver operator characteristics were used to report the sensitivity and specificity. Peptides differentiating controls from IBD originate from secreted phosphoprotein 24 (SPP24, p = 0.000086, 0.009); whereas those in remission and healthy can be differentiated in UC by SPP24 (p = 0.00023, 0.001), α-1-microglobulin (AMBP, p = 0.006) and CD by SPP24 (p = 0.019, 0.05). UC and CD can be differentiated by Guanylin (GUC2A, p = 0.001), and Secretogranin-1 (CHGB p = 0.035). Active and quiescent disease can also be differentiated in UC and CD by CHGB (p ≤ 0.023) SPP24 (p ≤ 0.023) and AMBP (UC p = 0.046). Five peptides discriminating IBD activity and severity had very little-to-no correlation to erythrocyte sedimentation rate, C-reactive protein, white cell or platelet counts. Three of these peptides were found to be binding partners to SPP24 protein alongside other known matrix proteins. These proteins have the potential to improve diagnosis and evaluate IBD activity, reducing the need for more invasive techniques. Data are available via ProteomeXchange with identifier PXD002821.
保护性肠道屏障的破坏会将效应分子和降解产物释放到血流中，使得血清和血浆成为理想的诊断介质。富含低质量蛋白质组作为诊断和监测炎症性肠病（IBD）活动的鉴别因子来源尚未被探索，这种方法比结肠镜检查侵入性更小。通过无标记定量质谱法评估了富含低质量血浆蛋白质组（<25 kDa）的差异。一组候选标志物进入了验证阶段以及“二级”美国食品药品监督管理局（FDA）级别的验证定量分析。通过多反应监测对血浆和血清中在上皮界面维持肠道屏障功能和稳态方面重要的蛋白质进行了定量，包括炎症性疾病（类风湿性关节炎对照）和非炎症性健康对照、溃疡性结肠炎（UC）以及克罗恩病（CD）患者。免疫印迹检测证实了这些蛋白质在血浆中的存在。使用相关性分析和受试者工作特征曲线来报告敏感性和特异性。区分对照与IBD的肽源自分泌磷蛋白24（SPP24，p = 0.000086，0.009）；而在UC中，缓解期和健康状态的个体可通过SPP24（p = 0.00023，0.001）、α-1-微球蛋白（AMBP，p = 0.006）进行区分，在CD中可通过SPP24（p = 0.019，0.05）进行区分。UC和CD可通过鸟苷素（GUC2A，p = 0.001）和分泌粒蛋白-1（CHGB，p = 0.035）进行区分。UC和CD中的活动期和静止期疾病也可通过CHGB（p≤0.023）、SPP24（p≤0.023）和AMBP（UC中p = 0.046）进行区分。区分IBD活动和严重程度的五种肽与红细胞沉降率、C反应蛋白、白细胞或血小板计数几乎没有或没有相关性。发现其中三种肽是SPP24蛋白以及其他已知基质蛋白的结合伴侣。这些蛋白质有潜力改善IBD的诊断并评估其活动情况，减少对侵入性更强技术的需求。数据可通过ProteomeXchange获取，标识符为PXD002821。
