Serum protein biomarkers for predicting type 1 diabetes

项目摘要/摘要 临床1型糖尿病(T1D)之前会出现疾病的无症状阶段，这很容易发生 以血清胰岛自身抗体(AAB)鉴定。然而，T1D领域面临着缺乏强有力的 预测标记物，特别是β细胞特异性血清标记物，可以准确地预测和揭示 T1D进展。这些标记对于开发新的、有效的干预措施至关重要 T1D。因此，该应用的总体目标是建立β细胞特异性血清蛋白质组 可以预测T1D免疫原性进展的信号。我们的具体假设是，正在进行的 在T1D前阶段对β细胞的自身免疫破坏将β细胞特异性蛋白特征转移到 血流，这些循环特征是评估β细胞质量的合适生物标记物，预测 β细胞破坏的轨迹，从而进入T1D的发育阶段。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全面识别 推测的β细胞特异性蛋白、异构体和翻译后修饰(PTM)来自胰岛和 纸巾。然后在从大规模收集的血清样本中对候选蛋白质进行优先排序和验证 纵向临床研究，如糖尿病预防试验(DPT-1) 超灵敏的靶向蛋白质组学技术。一组胰岛特异性蛋白的可用性(已经 通过我们的初步研究确认)，独特的超灵敏技术为我们提供了一流的 为T1D的发展建立新的预测血清标志物的机会。在我们的实验计划中，目标1 将专注于通过比较发现推测的β细胞特有的蛋白质、异构体和PTM T1D前与对照组胰腺和胰岛比较。AIM 2将对前100-200名进行初步筛选 基于它们在人血清中的可检测性以及它们的显著差异的血清 新发T1D受试者与对照组的血药浓度。AIM 3将对大多数 在DPT-1队列纵向收集的血清样本中，有希望的候选样本有10-30个。这个 比较将包括三组受试者：1)患有多种AAB并进展为 T1D，2)有多个AAB但未进展到T1D的T1D亲属，3)AAB阴性对照。我们 预计这一应用将使我们第一次能够成功地开发一个β细胞特异性的面板 血清标志物对预测T1D进展、评估β细胞具有深远的临床意义 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.