Serum protein biomarkers for predicting type 1 diabetes

项目总结/摘要 临床1型糖尿病（T1 D）之前是疾病的无症状阶段，其可以容易地被诊断为糖尿病。 通过血清胰岛自身抗体（AAb）鉴定。然而，T1 D领域受到缺乏有效的 预测性标志物，特别是β细胞特异性血清标志物，其可以准确预测和揭示 T1 D进展。这些标志物对于开发新的有效干预措施至关重要， T1D因此，本申请的总体目标是建立β细胞特异性血清蛋白质组学。 可以预测T1 D免疫原性进展的特征。我们的假设是， 在前T1 D阶段，β细胞的自身免疫性破坏将β细胞特异性蛋白质特征释放到细胞中。 血流，并且这些循环特征是用于评估β细胞质量、预测 β细胞破坏的轨迹以及T1 D的发展阶段。为了实现这一目标， 我们的基本原理是应用一种以假设为中心的综合发现方法， 来自胰岛的推定的β细胞特异性蛋白质、同种型和翻译后修饰（PTM）， 组织中然后，在从大规模免疫组织化学中收集的血清样品中对候选蛋白进行优先级排序和验证。 纵向临床研究，如糖尿病预防试验（DPT-1），通过实施 超灵敏的靶向蛋白质组学技术一组胰岛特异性蛋白质的可用性（已经 通过我们的初步研究确定）和独特的超灵敏技术为我们提供了一流的 有机会建立T1 D发展的新预测血清标志物。在我们的实验计划中，目标1 将集中于综合发现推定的β细胞特异性蛋白质，异构体，和PTM通过比较 T1 D前与对照胰腺和胰岛。Aim 2将对前100-200名进行初步筛选 候选物基于它们在人血清中的可检测性以及它们的显著差异血清 在新发T1 D受试者与对照中的浓度。Aim 3将对大多数 在DPT-1队列中纵向收集的血清样品中有希望有10-30个候选者。的 比较将包括三组受试者：1）T1 D亲属，他们有多种AAb，并进展到 T1 D，2）具有多种AAb但未进展为T1 D的T1 D亲属，和3）AAb阴性对照。我们 预计这项应用将使我们能够首次成功开发一组β细胞特异性 血清标志物，这将有深远的临床意义，预测T1 D进展，评估β细胞 质量，并监测治疗效果。

PROJECT SUMMARY/ABSTRACT Clinical type 1 diabetes (T1D) is preceded by an asymptomatic phase of the disease, which can be readily identified by serum islet autoantibodies(AAb). However, the T1D field is challenged by the lack of potent predictive markers, particularly β-cell specific serum markers that can accurately predict and reveal the rate of T1D progression. The markers of such are critical in the development of novel and effective interventions for T1D. Therefore, the overall objective of this application is to establish β-cell specific serum proteomic signatures that can predict the immunogenic progression of T1D. Our specific hypothesis is that the ongoing autoimmune destruction of β-cells during the pre-T1D phase sheds β-cell-specific protein signatures into the blood stream, and these circulatory signatures are suitable biomarkers for assessing β-cell mass, predicting the trajectory of β-cell destruction and thus the developmental stage of T1D. To pursue this objective, our rationale is to apply a hypothesis-focused integrative discovery approach to comprehensively identify putative β-cell specific proteins, isoforms, and posttranslational modifications (PTMs) from pancreatic islets and tissues. Protein candidates are then prioritized and validated in the serum samples collected from large-scale longitudinal clinical studies such as the Diabetes Prevention Trial (DPT-1) via the implementation of an ultrasensitive targeted proteomic technology. The availability of a set of islet-specific proteins (already identified through our preliminary studies) and the unique ultrasensitive technology provides us a first-rate opportunity to establish novel predictive serum markers for T1D development. In our experimental plan, Aim 1 will focus on the integrative discovery of putative β-cell specific proteins, isoforms, and PTMs by comparing pre-T1D versus control pancreas and islets. Aim 2 will conduct an initial screening for the top 100-200 candidates based on their detectability in human serum as well as their significant differential serum concentration in new-onset T1D subjects versus controls. Aim 3 will perform targeted validation of the most promising 10-30 candidates in the serum samples longitudinally collected in the DPT-1 cohort. The comparison will include three groups of subjects: 1) T1D relatives who had multiple AAb and progressed to T1D 2) T1D relatives who had multiple AAb but did not progress to T1D, and 3) AAb negative controls. We anticipate this application will enable us for the first time to successfully develop a panel of β-cell specific serum markers, which will have a profound clinical implication for predicting T1D progression, assessing β-cell mass, and monitoring the treatment efficacy.