Protein Phenotyping of SLE Nephritis Flare Cycle

描述（由申请人提供）：人类SLE累及肾脏是常见的，通常是严重的。治疗通常是有效的，但伴随着相当大的毒性。如果能够及早开始治疗，根据疾病的严重程度量身定制，并在注定会有持久反应的患者中迅速逐渐减少治疗，那么治疗的发病率可以大大降低。以这种方式使用治疗需要生物标志物来预测SLE肾脏耀斑的发作、耀斑的严重程度和对治疗的反应。这个试点项目的目的是确定是否可以通过检查整个肾脏耀斑周期的尿蛋白质组来模拟SLE肾脏耀斑的表型。这项工作将利用俄亥俄州立大学开发的俄亥俄系统性红斑狼疮研究数据库和标本库，其中包含一系列临床信息和来自系统性红斑狼疮患者队列的尿液和血浆样本，这些患者每2个月前瞻性随访5年。Aim 1将使用表面增强激光解吸/电离飞行时间质谱法（SELDI-TOF-MS）来测试分析SLE患者在肾脏耀斑发作前、耀斑发作期间和耀斑治疗后尿液蛋白质组变化的可行性。在肾脏耀斑发生前改变表达的蛋白可能预示着即将发生的耀斑和/或耀斑严重程度。在耀斑期间差异表达的蛋白质可能与肾损伤的发病机制有关，因此可能反映潜在的新的治疗靶点。耀斑治疗后蛋白质的变化可能有助于预测对治疗的反应。Aim 2将测试使用液相色谱-串联质谱法对Aim 1中SELDI-TOF-MS检测到的差异表达蛋白进行阳性鉴定的可行性。在Aim 3中，在Aim 2中确定的候选SLE肾炎生物标志物将在原始样本集中使用免疫检测或基于定量质谱的技术进行量化。总之，该项目有望通过监测SLE肾炎发作周期尿液蛋白质组的变化来证明生物标志物鉴定和验证的可行性。公共卫生相关性：这项研究将促进新型诊断工具的发展，以预测SLE肾耀斑的发病、严重程度和结局。这些工具将基于SLE肾脏耀斑期间尿蛋白的变化。这将导致早期诊断和治疗，从而更有效地使用现有药物，从而改善SLE肾炎的临床结果。

DESCRIPTION (provided by applicant): Kidney involvement in human SLE is common and usually severe. Treatment is often effective, but accompanied by considerable toxicity. Morbidity from treatment could be considerably reduced if it could be started early, tailored to disease severity, and tapered off quickly in patients destined to have a durable response. To use therapy in this way requires biomarkers that predict the onset of SLE renal flare, the severity of the flare, and the response to therapy. The goal of this pilot project is to determine whether the phenotype of SLE renal flare can be modeled by examining the urine proteome throughout the renal flare cycle. This work will utilize the Ohio SLE Study database and specimen bank, developed at the Ohio State University, and which contains serial clinical information and urine and plasma samples from a cohort of SLE patients followed prospectively every 2 months over 5 years. Aim 1 will use surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) to test the feasibility of profiling changes in the urine proteome of SLE patients before renal flare onset, during flare, and following treatment of flare. Proteins that change expression before a renal flare may be forecasters of impending flare and/or flare severity. Proteins that are differentially expressed during flare may be relevant to the pathogenesis of kidney injury, and may thus reflect potential novel therapeutic targets. Proteins that change after flare treatment may be useful in predicting response to treatment. Aim 2 will test the feasibility of using liquid chromatography-tandem mass spectrometry to positively identify differentially expressed proteins detected by SELDI-TOF-MS in Aim 1. In Aim 3, candidate SLE nephritis biomarkers identified in Aim 2 will be quantified in the original sample set using immunodetection or quantitative mass spectrometery-based techniques. In summary, this project is expected to demonstrate the feasibility of biomarker identification and validation by monitoring changes in the urine proteome during the SLE nephritis flare cycle. PUBLIC HEALTH RELEVANCE: This research will facilitate the development of novel diagnostic tools to predict onset, severity, and outcome of SLE renal flare. These tools will be based on changes in urine proteins during SLE renal flares. This will lead to earlier diagnosis and treatment, and thus more effective use of currently available medications, resulting in improved clinical outcomes for SLE nephritis.