Validation of Novel Peptide/Protein Markers for Diagnosis of Type 1 Diabetes

描述(申请人提供)：1型糖尿病(T1D)是由遗传易感个体中环境因素的复杂相互作用引发的产生胰岛素的胰腺细胞的自身免疫破坏所致。本实验室以前开展的研究揭示了一组新的多肽和相应的蛋白质具有很好的T1D与健康对照的区分能力，使用基于液质联用(LC-MS)的蛋白质组学分析来自糖尿病抗体标准化计划队列的人血清和血浆样本，样本量有限。该项目的长期目标是为临床干预T1D提供充分有效的诊断和预后标志物，并了解T1D的发病机制，从而最终预防T1D。在目前的应用中，以青年糖尿病自身免疫研究(DAISY)为T1D队列的父代，我们将使用我们的多重高效液相-多反应监测-质谱仪(LC-MRM-MS)平台和最先进的完整蛋白质分离与高分辨率串联质谱仪相结合的技术，从独立的大规模T1D队列以及患有与T1D相似临床和免疫学结果的疾病患者的大量(n=2090)血清/血浆中检测这些生物标志物。具体地说，我们提出了以下目标：目的1，建立一个独立的T1D队列中标记多肽的敏感性，并提供这些多肽在诊断T1D中的准确阈值；目的2，利用2型糖尿病、乳糜泻和炎症性肠病的疾病对照，建立标记多肽的特异性；目的3，鉴定关键标记多肽的蛋白质异构体，在这种疾病的发病机制中具有潜在的作用；目的4，验证大规模临床T1D队列中的多肽生物标记物的有效性；以及目的5，利用菊花队列的纵向样本，建立有效的多肽生物标记物的预后价值。本研究的结果将证实这些肽/蛋白标记物在诊断T1D方面的实用性和特异性，进一步了解该病的发病机制，并为T1D的预后、干预和预防提供新的策略基础。

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) results from autoimmune destruction of insulin-producing pancreatic ¿-cells triggered by complex interactions of environmental factors in genetically predisposed individuals. Previous studies carried out in this laboratory revealed a panel of novel peptides and corresponding proteins having very good discriminating power of T1D from healthy controls using liquid chromatography-mass spectrometry (LC-MS) based proteomics analyses of human blood serum and plasma samples from a Diabetes Antibody Standardization Program cohort with limited sample size. The long-term goal of this project is to provide thoroughly validated diagnostic and prognostic markers to the clinical community for intervention of T1D, and to understand the pathogenesis of T1D for its ultimate prevention. In the present application, using The Diabetes Autoimmunity in the Young Study (DAISY) as a parent T1D cohort, we will use our multiplexed liquid chromatography- multiple reaction monitoring-mass spectrometry (LC-MRM-MS) platform and state-of-the-art technologies on intact protein separation coupled with high resolution tandem mass spectrometry to measure these biomarkers in large numbers (n = 2090) of blood serum/plasma from independent, large scale T1D cohorts, as well as from individuals having diseases that share similar clinical and immunological outcomes with T1D. Specifically, we propose the following aims: Aim 1, to establish the sensitivity of marker peptides in an independent T1D cohort and provide accurate threshold values of these peptides in diagnosis of T1D; Aim 2, to establish the specificity of marker peptides using disease controls of type 2 diabetes, celiac disease and inflammatory bowel disease; Aim 3, to identify the protein isoforms of key marker peptides having potential roles in pathogenesis of this disease; Aim 4, to validate the peptide biomarkers in large scale clinical T1D cohorts blinded to the investigators; and Aim 5, to establish the prognostic value of validated peptide biomarkers with longitudinal samples from the DAISY cohort. The outcome of the proposed research will confirm the utility and specificity of these peptide/protein markers for diagnosing T1D, gain further insight to the pathogenesis of this disease, and provide foundations for new strategies in T1D prognosis, intervention and prevention.